This document provides an overview of advanced molecular biology techniques including recombinant DNA technologies, gene therapy, genetic modification of plants and animals, and tools used in genetic engineering. Key techniques discussed include using restriction enzymes and DNA ligase to create recombinant DNA, gel electrophoresis to separate DNA fragments, PCR to make copies of DNA, and bacterial plasmids to clone genes. Strategies for genetically engineering bacteria and applications of gene therapy like treating immune deficiencies are also summarized.
PRINCIPLES OF PLANT BIOTECHNOLOGY
Subham Mandal ( Student )
B.Sc Horticulture , 2nd year
Uttar Banga Krishi Viswavidyalaya
Disclaimer : I am also a student so.. read it at your own risk
SUMMARY :
- Gene Transfer:
1. Agrobacterium-mediated transformation
2. Biolistic or particle bombardment
3. Electroporation
4. Microinjection
5. Protoplast fusion
- Procedure of Gene Cloning:
1. Isolation of DNA
2. Preparation of vector
3. Insertion of DNA
4. Transformation
5. Identification/screening
- PCR:
1. Denaturation
2. Annealing
3. Extension
- DNA fingerprinting:
1. DNA extraction
2. DNA fragmentation
3. Gel electrophoresis
4. Southern blotting
5. Hybridization
6. Detection
7. Analysis
- Transgenic:
1. Bt Cotton
2. Bt Brinjal
3. Golden Rice
4. Bt Rice
5. GM Mustard
- Molecular markers:
1. RFLP
2. AFLP
3. SSR
4. SNP
5. Indels
- Vectors:
1. Plasmid vectors
2. Cosmid vectors
3. Bacterial artificial chromosome (BAC) vector
- MAS (Marker-Assisted Selection):
1. Improvement of yield and quality
2. Enhancement of nutritional content
3. Development of stress-tolerant crops
4. Identification of disease-resistant plants
5. Improvement of crop traits through genetic modification
Anther Culture: Culturing immature pollen grains to produce haploid plantlets for breeding and genetic research.
Embryo Culture: Growing and developing plant embryos in vitro for clonal propagation and study of embryogenesis.
Pollen Culture: Culturing mature pollen grains to produce haploid plantlets and create new cultivars.
Ovule Culture: Culturing ovules for haploid or doubled haploid plant production and hybridization.
Somatic Embryogenesis: Inducing embryonic structures from somatic cells for clonal propagation and genetic modification.
Meristem Culture: Culturing the apical meristem for virus-free stock recovery and micropropagation.
This document discusses genetic engineering tools and techniques. It explains that genetic engineers use restriction enzymes, plasmids, and promoters to insert genes from one organism into another. The inserted genes are expressed to produce proteins like insulin. Gel electrophoresis and fluorescent markers are described as tools to analyze DNA and identify transformed bacteria. The overall aim is to manipulate genes between species to study and modify organisms.
Roughly based on Chapter 11 Biotechnology: Principles and Processes and Chapter 12 Biotechnology and its Applications of Class 12 NCERT for final brush-up before the exams
DNA cloning is the process of making multiple, identical copies of a particular piece of DNA. In a typical DNA cloning procedure, the gene or other DNA fragment of interest (perhaps a gene for a medically important human protein) is first inserted into a circular piece of DNA called a plasmid.- [https://www.khanacademy.org/science/...dna.../dna-cloning.../a/overview-dna-cloning]
This document provides an overview of various gene transfer tools and techniques. It discusses vector-mediated methods like Agrobacterium and viral vectors as well as direct or vector-less methods such as electroporation, biolistics, microinjection, liposome mediated, and calcium phosphate mediated gene transfer. For each method, it describes the basic process and provides some key details and applications. It also notes some advantages and limitations of different techniques. The document aims to inform readers about the various options available for inserting genes into plant cells.
Recombinant DNA (rDNA) refers to DNA created outside living cells by joining DNA from multiple sources. Common techniques for creating rDNA include restriction enzymes to cut DNA strands, ligation to join strands, and transformation or transfection to introduce rDNA into host cells. Vectors like plasmids, viruses, and artificial chromosomes are often used to replicate and express rDNA in host cells. rDNA techniques have applications in gene cloning, DNA sequencing, genetic engineering of plants and animals, and gene therapy to treat diseases.
DNA Transfection in Animal tissue culture and its methods.pptxMethusharma
You will learn the definition of DNA transfection in this presentation its examples, along with the procedures that are employed, through the use of organised flowcharts and diagrams. The Animal Biotechnology course, it is the first technique to learn.
Gene cloning allows for the creation of identical copies of genes. It involves amplifying genes using polymerase chain reaction, cutting DNA with restriction enzymes, and joining DNA fragments together with DNA ligase. Cloning vectors like plasmids and bacteriophages are used to move genes into host cells. Transformed cells are selected using antibiotic resistance or reporter genes. Cloned genes have applications in pharmaceutical production, disease diagnosis using probes or PCR, and controlling insect pests by producing bacterial pesticides, transgenic plants, or viral pesticides.
PRINCIPLES OF PLANT BIOTECHNOLOGY
Subham Mandal ( Student )
B.Sc Horticulture , 2nd year
Uttar Banga Krishi Viswavidyalaya
Disclaimer : I am also a student so.. read it at your own risk
SUMMARY :
- Gene Transfer:
1. Agrobacterium-mediated transformation
2. Biolistic or particle bombardment
3. Electroporation
4. Microinjection
5. Protoplast fusion
- Procedure of Gene Cloning:
1. Isolation of DNA
2. Preparation of vector
3. Insertion of DNA
4. Transformation
5. Identification/screening
- PCR:
1. Denaturation
2. Annealing
3. Extension
- DNA fingerprinting:
1. DNA extraction
2. DNA fragmentation
3. Gel electrophoresis
4. Southern blotting
5. Hybridization
6. Detection
7. Analysis
- Transgenic:
1. Bt Cotton
2. Bt Brinjal
3. Golden Rice
4. Bt Rice
5. GM Mustard
- Molecular markers:
1. RFLP
2. AFLP
3. SSR
4. SNP
5. Indels
- Vectors:
1. Plasmid vectors
2. Cosmid vectors
3. Bacterial artificial chromosome (BAC) vector
- MAS (Marker-Assisted Selection):
1. Improvement of yield and quality
2. Enhancement of nutritional content
3. Development of stress-tolerant crops
4. Identification of disease-resistant plants
5. Improvement of crop traits through genetic modification
Anther Culture: Culturing immature pollen grains to produce haploid plantlets for breeding and genetic research.
Embryo Culture: Growing and developing plant embryos in vitro for clonal propagation and study of embryogenesis.
Pollen Culture: Culturing mature pollen grains to produce haploid plantlets and create new cultivars.
Ovule Culture: Culturing ovules for haploid or doubled haploid plant production and hybridization.
Somatic Embryogenesis: Inducing embryonic structures from somatic cells for clonal propagation and genetic modification.
Meristem Culture: Culturing the apical meristem for virus-free stock recovery and micropropagation.
This document discusses genetic engineering tools and techniques. It explains that genetic engineers use restriction enzymes, plasmids, and promoters to insert genes from one organism into another. The inserted genes are expressed to produce proteins like insulin. Gel electrophoresis and fluorescent markers are described as tools to analyze DNA and identify transformed bacteria. The overall aim is to manipulate genes between species to study and modify organisms.
Roughly based on Chapter 11 Biotechnology: Principles and Processes and Chapter 12 Biotechnology and its Applications of Class 12 NCERT for final brush-up before the exams
DNA cloning is the process of making multiple, identical copies of a particular piece of DNA. In a typical DNA cloning procedure, the gene or other DNA fragment of interest (perhaps a gene for a medically important human protein) is first inserted into a circular piece of DNA called a plasmid.- [https://www.khanacademy.org/science/...dna.../dna-cloning.../a/overview-dna-cloning]
This document provides an overview of various gene transfer tools and techniques. It discusses vector-mediated methods like Agrobacterium and viral vectors as well as direct or vector-less methods such as electroporation, biolistics, microinjection, liposome mediated, and calcium phosphate mediated gene transfer. For each method, it describes the basic process and provides some key details and applications. It also notes some advantages and limitations of different techniques. The document aims to inform readers about the various options available for inserting genes into plant cells.
Recombinant DNA (rDNA) refers to DNA created outside living cells by joining DNA from multiple sources. Common techniques for creating rDNA include restriction enzymes to cut DNA strands, ligation to join strands, and transformation or transfection to introduce rDNA into host cells. Vectors like plasmids, viruses, and artificial chromosomes are often used to replicate and express rDNA in host cells. rDNA techniques have applications in gene cloning, DNA sequencing, genetic engineering of plants and animals, and gene therapy to treat diseases.
DNA Transfection in Animal tissue culture and its methods.pptxMethusharma
You will learn the definition of DNA transfection in this presentation its examples, along with the procedures that are employed, through the use of organised flowcharts and diagrams. The Animal Biotechnology course, it is the first technique to learn.
Gene cloning allows for the creation of identical copies of genes. It involves amplifying genes using polymerase chain reaction, cutting DNA with restriction enzymes, and joining DNA fragments together with DNA ligase. Cloning vectors like plasmids and bacteriophages are used to move genes into host cells. Transformed cells are selected using antibiotic resistance or reporter genes. Cloned genes have applications in pharmaceutical production, disease diagnosis using probes or PCR, and controlling insect pests by producing bacterial pesticides, transgenic plants, or viral pesticides.
DNA technology can be used to identify genes for specific traits, transfer genes between organisms, cure diseases and treat genetic disorders. Key tools for manipulating DNA include restriction enzymes, gel electrophoresis, DNA ligase and polymerase chain reaction. Genes can be extracted and inserted into plasmids in bacteria using these tools to produce proteins like insulin. While this technology has practical uses, it also raises ethical issues regarding privacy and environmental impact.
1. DNA technology uses restriction enzymes to cut DNA into segments that can be spliced together to transfer genes between organisms.
2. Genes for human insulin and other proteins can be cloned in bacteria to produce large quantities for medical use.
3. The human genome project aims to sequence all human DNA to better understand genes and lead to treatments for genetic disorders.
1. DNA technology uses restriction enzymes to cut DNA into segments that can be spliced together to clone genes. Restriction enzymes recognize specific DNA sequences and cut the DNA.
2. Cloning vectors like plasmids are used to transfer genes between organisms. A donor gene is inserted into the plasmid which is then placed into a host cell to replicate the gene.
3. Recombinant DNA combines DNA from different sources, like inserting a human gene for insulin into bacterial DNA. The bacteria then produces insulin protein in large quantities to treat diabetes.
Gene cloning involves isolating a specific gene from an organism's DNA and copying it. This is done by cutting the DNA into fragments using restriction enzymes, inserting the gene fragments into bacterial plasmids, and transforming the plasmids into bacteria. The transformed bacteria are cultured to form a DNA library containing many copies of the cloned gene. The library is screened to identify the bacteria containing the gene of interest. cDNA libraries are also created using reverse transcriptase to copy mRNA into DNA that can be expressed in bacteria without introns. Gene cloning is important for research but raises some ethical issues regarding antibiotic resistance and ownership of genetic information.
Gene Cloning Very Detailed Antibiotic Resistanceallyjer
Gene cloning involves isolating a specific gene from an organism's DNA and copying it. This is done by cutting the DNA into fragments using restriction enzymes, inserting the gene fragments into bacterial plasmids, and inserting the plasmids into bacteria through a process called transformation. The transformed bacteria are then grown into a DNA library, where each bacterial colony contains copies of the same DNA fragment. The library is screened to identify the colonies containing the gene of interest. cDNA libraries are also created using reverse transcriptase to copy mRNA into DNA that can be expressed in bacteria without introns. Gene cloning is done to study genes and their functions, identify mutations, and engineer organisms for useful purposes, but it raises some ethical issues regarding antibiotic resistance and
Gene cloning involves isolating a specific gene from an organism's DNA and copying it. This is done by cutting the DNA into fragments using restriction enzymes, inserting the gene fragments into bacterial plasmids, and inserting the plasmids into bacteria through a process called transformation. The transformed bacteria are then grown into a DNA library, where each bacterial colony contains copies of the inserted gene. The library is screened to identify the colonies containing the desired gene. cDNA libraries are also created using reverse transcriptase to copy mRNA into DNA without introns, allowing eukaryotic genes to be expressed in bacteria. Gene cloning is done to study genes and their functions, identify mutations, and engineer organisms for useful purposes, but it raises some ethical issues
Gene cloning involves isolating a specific gene from an organism's DNA and copying it. This is done by cutting the DNA into fragments using restriction enzymes, inserting the gene fragments into bacterial plasmids, and inserting the plasmids into bacteria through a process called transformation. The transformed bacteria are then grown into a DNA library containing many copies of the cloned gene. DNA libraries are screened to identify the bacteria containing the gene of interest. cDNA libraries are also created using reverse transcriptase to clone eukaryotic genes for expression in bacteria by removing introns. Gene cloning raises some ethical issues regarding antibiotic resistance genes and ownership of cloned genes.
Genetic engineering and Recombinant DNAHala AbuZied
Genetic engineering involves altering the DNA of living organisms using biotechnology. It includes techniques like changing single DNA base pairs, deleting or adding genes, or combining DNA from different species. Recombinant DNA technology is used to create recombinant DNA molecules by manipulating DNA in vitro and introducing them into host organisms. This allows bacteria to be engineered to produce human insulin through inserting the human insulin gene into bacterial plasmids. Genomic libraries can be created by ligating fragmented genomic or cDNA into plasmid vectors to transform bacteria and clone the entire genome.
Recombinant DNA (rDNA) technology involves combining DNA molecules from different sources into new combinations. DNA from one organism is cut and joined with DNA from another organism, and the resulting recombinant DNA is inserted into a host cell. This allows genes and DNA fragments to be expressed, amplified, and mass produced. The basic steps are isolation of genetic material, restriction enzyme digestion, amplification via PCR, ligation, insertion into a host cell, and isolation of recombinant cells. rDNA technology has many applications including basic research, production of therapeutic proteins, agriculture, medicine, and industry. Hybridoma technology uses cell fusion to produce monoclonal antibodies that are identical clones of a single parent cell.
recombinant dna tech_molecular genetics lect 2nd yr mt-1st semester.pdfBekarEmail
Recombinant DNA technology involves combining DNA sequences from different species that would not normally occur together. It allows for the large-scale production of functional proteins by cloning DNA fragments into plasmids, which are then inserted into host cells to generate multiple recombinant clones. Some applications of recombinant DNA technology include producing insulin, vaccines, fluorescent proteins, and pesticide-resistant crops.
This document provides an overview of plasmids. It begins by noting that plasmids were first introduced by American molecular biologist Joshua Lederberg in 1952. It then discusses that recombinant DNA technology involves inserting a gene of interest into a plasmid vector. Plasmids are described as circular DNA molecules that can replicate independently of the bacterial chromosome and confer selective advantages to bacteria. The document outlines the components and functions of plasmids, including origins of replication, antibiotic resistance genes, and multiple cloning sites. It also discusses the applications of plasmids in DNA cloning, genetic engineering, and production of therapeutic proteins.
Genetic engineering principle, tools, techniques, types and applicationTarun Kapoor
Basic principles of genetic engineering.
Study of cloning vectors, restriction endonucleases and DNA ligase.
Recombinant DNA technology. Application of genetic engineering in medicine.
Application of r DNA technology and genetic engineering in the products:
a. Interferon
b. Vaccines- hepatitis- B
c. Hormones- Insulin.
Polymerase chain reaction
Brief introduction to PCR
Basic principles of PCR
This document provides an overview of recombinant DNA technology. It begins by describing the basic components and structure of DNA, including nucleotides, nitrogen bases, and how DNA encodes genetic instructions. It then defines what a gene is and explains that recombinant DNA technology involves joining DNA fragments from different sources. The key steps are described as isolating the gene of interest, inserting it into a vector like a plasmid, introducing the vector into a host cell, and amplifying the recombinant DNA. A variety of applications are mentioned, such as producing pharmaceuticals, genetically modifying crops, and using bacteria to break down environmental waste.
Recombinant DNA technology allows DNA from different species to be isolated, cut, and spliced together to form new recombinant molecules. Key tools for recombinant DNA technology include restriction enzymes, ligases, polymerases, vectors, and host cells. Recombinant DNA technology has many applications, including producing human insulin and other proteins for medical use, genetically engineering plants for crop improvement, and DNA fingerprinting for criminal investigations.
Cloning vectors are small DNA molecules that are used to artificially carry foreign genetic material into host cells. They contain features like an origin of replication, antibiotic resistance genes, and restriction enzyme sites. The document discusses different types of cloning vectors used for plant gene cloning, including plasmids, Ti/Ri plasmids from Agrobacterium, and plant viruses. Agrobacterium-mediated transformation uses disarmed Ti plasmids from Agrobacterium tumefaciens and involves co-cultivation of plant explants with Agrobacterium, selection of transformed cells, and regeneration of whole plants. Binary vector systems are now commonly used, involving transfer of a binary plasmid without integration into the Ti plasmid
Biotechnology uses living organisms to produce useful materials. It has both traditional and innovative forms. Recombinant DNA technology allows genes to be transferred between organisms. This has led to important medical advances like producing insulin in bacteria. However, there are also concerns about GMOs, including possible allergic reactions in humans and the risk of transgenic genes escaping into the wild. Overall, biotechnology presents both opportunities and risks that require careful consideration.
This document discusses various methods for the production of pharmaceutical drugs through microbial biotechnology and genetic engineering. It covers topics like the production of antibiotics like penicillin and chlortetracycline through fermentation, the use of recombinant DNA technology to produce drugs in microorganisms, and the application of genetic manipulation techniques. It also summarizes strategies for producing drugs like human insulin through the transformation of E. coli or B. subtilis with human genes.
This document provides an overview of DNA cloning including:
1. The basic steps in DNA cloning including isolation of vector and gene source DNA, insertion into the vector, and introduction into cells.
2. Uses of polymerase chain reaction and restriction enzymes in cloning.
3. Applications of cloning such as recombinant protein production, genetically modified organisms, DNA fingerprinting, and gene therapy.
Bacterial plasmids are small, circular, extrachromosomal DNA molecules that are able to replicate independently of the bacterial chromosome. Plasmids are commonly found in bacteria and can carry genes conferring traits such as antibiotic resistance. Plasmids are useful genetic engineering tools as they can be used to insert foreign DNA and replicate this DNA within bacterial cells. Common plasmids include R plasmids containing antibiotic resistance genes and F plasmids involved in bacterial conjugation.
Recombinant DNA technology allows DNA from different species to be isolated, cut, spliced together, and replicated. This creates new "recombinant" DNA molecules. Key steps include using restriction enzymes to cut DNA into fragments, inserting fragments into cloning vectors like plasmids, and transforming host cells to replicate the recombinant DNA. PCR is also used to amplify specific DNA sequences. Recombinant DNA technology has many applications, including producing human proteins, diagnosing genetic diseases, and detecting bacteria and viruses.
DNA technology can be used to identify genes for specific traits, transfer genes between organisms, cure diseases and treat genetic disorders. Key tools for manipulating DNA include restriction enzymes, gel electrophoresis, DNA ligase and polymerase chain reaction. Genes can be extracted and inserted into plasmids in bacteria using these tools to produce proteins like insulin. While this technology has practical uses, it also raises ethical issues regarding privacy and environmental impact.
1. DNA technology uses restriction enzymes to cut DNA into segments that can be spliced together to transfer genes between organisms.
2. Genes for human insulin and other proteins can be cloned in bacteria to produce large quantities for medical use.
3. The human genome project aims to sequence all human DNA to better understand genes and lead to treatments for genetic disorders.
1. DNA technology uses restriction enzymes to cut DNA into segments that can be spliced together to clone genes. Restriction enzymes recognize specific DNA sequences and cut the DNA.
2. Cloning vectors like plasmids are used to transfer genes between organisms. A donor gene is inserted into the plasmid which is then placed into a host cell to replicate the gene.
3. Recombinant DNA combines DNA from different sources, like inserting a human gene for insulin into bacterial DNA. The bacteria then produces insulin protein in large quantities to treat diabetes.
Gene cloning involves isolating a specific gene from an organism's DNA and copying it. This is done by cutting the DNA into fragments using restriction enzymes, inserting the gene fragments into bacterial plasmids, and transforming the plasmids into bacteria. The transformed bacteria are cultured to form a DNA library containing many copies of the cloned gene. The library is screened to identify the bacteria containing the gene of interest. cDNA libraries are also created using reverse transcriptase to copy mRNA into DNA that can be expressed in bacteria without introns. Gene cloning is important for research but raises some ethical issues regarding antibiotic resistance and ownership of genetic information.
Gene Cloning Very Detailed Antibiotic Resistanceallyjer
Gene cloning involves isolating a specific gene from an organism's DNA and copying it. This is done by cutting the DNA into fragments using restriction enzymes, inserting the gene fragments into bacterial plasmids, and inserting the plasmids into bacteria through a process called transformation. The transformed bacteria are then grown into a DNA library, where each bacterial colony contains copies of the same DNA fragment. The library is screened to identify the colonies containing the gene of interest. cDNA libraries are also created using reverse transcriptase to copy mRNA into DNA that can be expressed in bacteria without introns. Gene cloning is done to study genes and their functions, identify mutations, and engineer organisms for useful purposes, but it raises some ethical issues regarding antibiotic resistance and
Gene cloning involves isolating a specific gene from an organism's DNA and copying it. This is done by cutting the DNA into fragments using restriction enzymes, inserting the gene fragments into bacterial plasmids, and inserting the plasmids into bacteria through a process called transformation. The transformed bacteria are then grown into a DNA library, where each bacterial colony contains copies of the inserted gene. The library is screened to identify the colonies containing the desired gene. cDNA libraries are also created using reverse transcriptase to copy mRNA into DNA without introns, allowing eukaryotic genes to be expressed in bacteria. Gene cloning is done to study genes and their functions, identify mutations, and engineer organisms for useful purposes, but it raises some ethical issues
Gene cloning involves isolating a specific gene from an organism's DNA and copying it. This is done by cutting the DNA into fragments using restriction enzymes, inserting the gene fragments into bacterial plasmids, and inserting the plasmids into bacteria through a process called transformation. The transformed bacteria are then grown into a DNA library containing many copies of the cloned gene. DNA libraries are screened to identify the bacteria containing the gene of interest. cDNA libraries are also created using reverse transcriptase to clone eukaryotic genes for expression in bacteria by removing introns. Gene cloning raises some ethical issues regarding antibiotic resistance genes and ownership of cloned genes.
Genetic engineering and Recombinant DNAHala AbuZied
Genetic engineering involves altering the DNA of living organisms using biotechnology. It includes techniques like changing single DNA base pairs, deleting or adding genes, or combining DNA from different species. Recombinant DNA technology is used to create recombinant DNA molecules by manipulating DNA in vitro and introducing them into host organisms. This allows bacteria to be engineered to produce human insulin through inserting the human insulin gene into bacterial plasmids. Genomic libraries can be created by ligating fragmented genomic or cDNA into plasmid vectors to transform bacteria and clone the entire genome.
Recombinant DNA (rDNA) technology involves combining DNA molecules from different sources into new combinations. DNA from one organism is cut and joined with DNA from another organism, and the resulting recombinant DNA is inserted into a host cell. This allows genes and DNA fragments to be expressed, amplified, and mass produced. The basic steps are isolation of genetic material, restriction enzyme digestion, amplification via PCR, ligation, insertion into a host cell, and isolation of recombinant cells. rDNA technology has many applications including basic research, production of therapeutic proteins, agriculture, medicine, and industry. Hybridoma technology uses cell fusion to produce monoclonal antibodies that are identical clones of a single parent cell.
recombinant dna tech_molecular genetics lect 2nd yr mt-1st semester.pdfBekarEmail
Recombinant DNA technology involves combining DNA sequences from different species that would not normally occur together. It allows for the large-scale production of functional proteins by cloning DNA fragments into plasmids, which are then inserted into host cells to generate multiple recombinant clones. Some applications of recombinant DNA technology include producing insulin, vaccines, fluorescent proteins, and pesticide-resistant crops.
This document provides an overview of plasmids. It begins by noting that plasmids were first introduced by American molecular biologist Joshua Lederberg in 1952. It then discusses that recombinant DNA technology involves inserting a gene of interest into a plasmid vector. Plasmids are described as circular DNA molecules that can replicate independently of the bacterial chromosome and confer selective advantages to bacteria. The document outlines the components and functions of plasmids, including origins of replication, antibiotic resistance genes, and multiple cloning sites. It also discusses the applications of plasmids in DNA cloning, genetic engineering, and production of therapeutic proteins.
Genetic engineering principle, tools, techniques, types and applicationTarun Kapoor
Basic principles of genetic engineering.
Study of cloning vectors, restriction endonucleases and DNA ligase.
Recombinant DNA technology. Application of genetic engineering in medicine.
Application of r DNA technology and genetic engineering in the products:
a. Interferon
b. Vaccines- hepatitis- B
c. Hormones- Insulin.
Polymerase chain reaction
Brief introduction to PCR
Basic principles of PCR
This document provides an overview of recombinant DNA technology. It begins by describing the basic components and structure of DNA, including nucleotides, nitrogen bases, and how DNA encodes genetic instructions. It then defines what a gene is and explains that recombinant DNA technology involves joining DNA fragments from different sources. The key steps are described as isolating the gene of interest, inserting it into a vector like a plasmid, introducing the vector into a host cell, and amplifying the recombinant DNA. A variety of applications are mentioned, such as producing pharmaceuticals, genetically modifying crops, and using bacteria to break down environmental waste.
Recombinant DNA technology allows DNA from different species to be isolated, cut, and spliced together to form new recombinant molecules. Key tools for recombinant DNA technology include restriction enzymes, ligases, polymerases, vectors, and host cells. Recombinant DNA technology has many applications, including producing human insulin and other proteins for medical use, genetically engineering plants for crop improvement, and DNA fingerprinting for criminal investigations.
Cloning vectors are small DNA molecules that are used to artificially carry foreign genetic material into host cells. They contain features like an origin of replication, antibiotic resistance genes, and restriction enzyme sites. The document discusses different types of cloning vectors used for plant gene cloning, including plasmids, Ti/Ri plasmids from Agrobacterium, and plant viruses. Agrobacterium-mediated transformation uses disarmed Ti plasmids from Agrobacterium tumefaciens and involves co-cultivation of plant explants with Agrobacterium, selection of transformed cells, and regeneration of whole plants. Binary vector systems are now commonly used, involving transfer of a binary plasmid without integration into the Ti plasmid
Biotechnology uses living organisms to produce useful materials. It has both traditional and innovative forms. Recombinant DNA technology allows genes to be transferred between organisms. This has led to important medical advances like producing insulin in bacteria. However, there are also concerns about GMOs, including possible allergic reactions in humans and the risk of transgenic genes escaping into the wild. Overall, biotechnology presents both opportunities and risks that require careful consideration.
This document discusses various methods for the production of pharmaceutical drugs through microbial biotechnology and genetic engineering. It covers topics like the production of antibiotics like penicillin and chlortetracycline through fermentation, the use of recombinant DNA technology to produce drugs in microorganisms, and the application of genetic manipulation techniques. It also summarizes strategies for producing drugs like human insulin through the transformation of E. coli or B. subtilis with human genes.
This document provides an overview of DNA cloning including:
1. The basic steps in DNA cloning including isolation of vector and gene source DNA, insertion into the vector, and introduction into cells.
2. Uses of polymerase chain reaction and restriction enzymes in cloning.
3. Applications of cloning such as recombinant protein production, genetically modified organisms, DNA fingerprinting, and gene therapy.
Bacterial plasmids are small, circular, extrachromosomal DNA molecules that are able to replicate independently of the bacterial chromosome. Plasmids are commonly found in bacteria and can carry genes conferring traits such as antibiotic resistance. Plasmids are useful genetic engineering tools as they can be used to insert foreign DNA and replicate this DNA within bacterial cells. Common plasmids include R plasmids containing antibiotic resistance genes and F plasmids involved in bacterial conjugation.
Recombinant DNA technology allows DNA from different species to be isolated, cut, spliced together, and replicated. This creates new "recombinant" DNA molecules. Key steps include using restriction enzymes to cut DNA into fragments, inserting fragments into cloning vectors like plasmids, and transforming host cells to replicate the recombinant DNA. PCR is also used to amplify specific DNA sequences. Recombinant DNA technology has many applications, including producing human proteins, diagnosing genetic diseases, and detecting bacteria and viruses.
Microbial interaction
Microorganisms interacts with each other and can be physically associated with another organisms in a variety of ways.
One organism can be located on the surface of another organism as an ectobiont or located within another organism as endobiont.
Microbial interaction may be positive such as mutualism, proto-cooperation, commensalism or may be negative such as parasitism, predation or competition
Types of microbial interaction
Positive interaction: mutualism, proto-cooperation, commensalism
Negative interaction: Ammensalism (antagonism), parasitism, predation, competition
I. Mutualism:
It is defined as the relationship in which each organism in interaction gets benefits from association. It is an obligatory relationship in which mutualist and host are metabolically dependent on each other.
Mutualistic relationship is very specific where one member of association cannot be replaced by another species.
Mutualism require close physical contact between interacting organisms.
Relationship of mutualism allows organisms to exist in habitat that could not occupied by either species alone.
Mutualistic relationship between organisms allows them to act as a single organism.
Examples of mutualism:
i. Lichens:
Lichens are excellent example of mutualism.
They are the association of specific fungi and certain genus of algae. In lichen, fungal partner is called mycobiont and algal partner is called
II. Syntrophism:
It is an association in which the growth of one organism either depends on or improved by the substrate provided by another organism.
In syntrophism both organism in association gets benefits.
Compound A
Utilized by population 1
Compound B
Utilized by population 2
Compound C
utilized by both Population 1+2
Products
In this theoretical example of syntrophism, population 1 is able to utilize and metabolize compound A, forming compound B but cannot metabolize beyond compound B without co-operation of population 2. Population 2is unable to utilize compound A but it can metabolize compound B forming compound C. Then both population 1 and 2 are able to carry out metabolic reaction which leads to formation of end product that neither population could produce alone.
Examples of syntrophism:
i. Methanogenic ecosystem in sludge digester
Methane produced by methanogenic bacteria depends upon interspecies hydrogen transfer by other fermentative bacteria.
Anaerobic fermentative bacteria generate CO2 and H2 utilizing carbohydrates which is then utilized by methanogenic bacteria (Methanobacter) to produce methane.
ii. Lactobacillus arobinosus and Enterococcus faecalis:
In the minimal media, Lactobacillus arobinosus and Enterococcus faecalis are able to grow together but not alone.
The synergistic relationship between E. faecalis and L. arobinosus occurs in which E. faecalis require folic acid
Evidence of Jet Activity from the Secondary Black Hole in the OJ 287 Binary S...Sérgio Sacani
Wereport the study of a huge optical intraday flare on 2021 November 12 at 2 a.m. UT in the blazar OJ287. In the binary black hole model, it is associated with an impact of the secondary black hole on the accretion disk of the primary. Our multifrequency observing campaign was set up to search for such a signature of the impact based on a prediction made 8 yr earlier. The first I-band results of the flare have already been reported by Kishore et al. (2024). Here we combine these data with our monitoring in the R-band. There is a big change in the R–I spectral index by 1.0 ±0.1 between the normal background and the flare, suggesting a new component of radiation. The polarization variation during the rise of the flare suggests the same. The limits on the source size place it most reasonably in the jet of the secondary BH. We then ask why we have not seen this phenomenon before. We show that OJ287 was never before observed with sufficient sensitivity on the night when the flare should have happened according to the binary model. We also study the probability that this flare is just an oversized example of intraday variability using the Krakow data set of intense monitoring between 2015 and 2023. We find that the occurrence of a flare of this size and rapidity is unlikely. In machine-readable Tables 1 and 2, we give the full orbit-linked historical light curve of OJ287 as well as the dense monitoring sample of Krakow.
Anti-Universe And Emergent Gravity and the Dark UniverseSérgio Sacani
Recent theoretical progress indicates that spacetime and gravity emerge together from the entanglement structure of an underlying microscopic theory. These ideas are best understood in Anti-de Sitter space, where they rely on the area law for entanglement entropy. The extension to de Sitter space requires taking into account the entropy and temperature associated with the cosmological horizon. Using insights from string theory, black hole physics and quantum information theory we argue that the positive dark energy leads to a thermal volume law contribution to the entropy that overtakes the area law precisely at the cosmological horizon. Due to the competition between area and volume law entanglement the microscopic de Sitter states do not thermalise at sub-Hubble scales: they exhibit memory effects in the form of an entropy displacement caused by matter. The emergent laws of gravity contain an additional ‘dark’ gravitational force describing the ‘elastic’ response due to the entropy displacement. We derive an estimate of the strength of this extra force in terms of the baryonic mass, Newton’s constant and the Hubble acceleration scale a0 = cH0, and provide evidence for the fact that this additional ‘dark gravity force’ explains the observed phenomena in galaxies and clusters currently attributed to dark matter.
Sexuality - Issues, Attitude and Behaviour - Applied Social Psychology - Psyc...PsychoTech Services
A proprietary approach developed by bringing together the best of learning theories from Psychology, design principles from the world of visualization, and pedagogical methods from over a decade of training experience, that enables you to: Learn better, faster!
Embracing Deep Variability For Reproducibility and Replicability
Abstract: Reproducibility (aka determinism in some cases) constitutes a fundamental aspect in various fields of computer science, such as floating-point computations in numerical analysis and simulation, concurrency models in parallelism, reproducible builds for third parties integration and packaging, and containerization for execution environments. These concepts, while pervasive across diverse concerns, often exhibit intricate inter-dependencies, making it challenging to achieve a comprehensive understanding. In this short and vision paper we delve into the application of software engineering techniques, specifically variability management, to systematically identify and explicit points of variability that may give rise to reproducibility issues (eg language, libraries, compiler, virtual machine, OS, environment variables, etc). The primary objectives are: i) gaining insights into the variability layers and their possible interactions, ii) capturing and documenting configurations for the sake of reproducibility, and iii) exploring diverse configurations to replicate, and hence validate and ensure the robustness of results. By adopting these methodologies, we aim to address the complexities associated with reproducibility and replicability in modern software systems and environments, facilitating a more comprehensive and nuanced perspective on these critical aspects.
https://hal.science/hal-04582287
SDSS1335+0728: The awakening of a ∼ 106M⊙ black hole⋆Sérgio Sacani
Context. The early-type galaxy SDSS J133519.91+072807.4 (hereafter SDSS1335+0728), which had exhibited no prior optical variations during the preceding two decades, began showing significant nuclear variability in the Zwicky Transient Facility (ZTF) alert stream from December 2019 (as ZTF19acnskyy). This variability behaviour, coupled with the host-galaxy properties, suggests that SDSS1335+0728 hosts a ∼ 106M⊙ black hole (BH) that is currently in the process of ‘turning on’. Aims. We present a multi-wavelength photometric analysis and spectroscopic follow-up performed with the aim of better understanding the origin of the nuclear variations detected in SDSS1335+0728. Methods. We used archival photometry (from WISE, 2MASS, SDSS, GALEX, eROSITA) and spectroscopic data (from SDSS and LAMOST) to study the state of SDSS1335+0728 prior to December 2019, and new observations from Swift, SOAR/Goodman, VLT/X-shooter, and Keck/LRIS taken after its turn-on to characterise its current state. We analysed the variability of SDSS1335+0728 in the X-ray/UV/optical/mid-infrared range, modelled its spectral energy distribution prior to and after December 2019, and studied the evolution of its UV/optical spectra. Results. From our multi-wavelength photometric analysis, we find that: (a) since 2021, the UV flux (from Swift/UVOT observations) is four times brighter than the flux reported by GALEX in 2004; (b) since June 2022, the mid-infrared flux has risen more than two times, and the W1−W2 WISE colour has become redder; and (c) since February 2024, the source has begun showing X-ray emission. From our spectroscopic follow-up, we see that (i) the narrow emission line ratios are now consistent with a more energetic ionising continuum; (ii) broad emission lines are not detected; and (iii) the [OIII] line increased its flux ∼ 3.6 years after the first ZTF alert, which implies a relatively compact narrow-line-emitting region. Conclusions. We conclude that the variations observed in SDSS1335+0728 could be either explained by a ∼ 106M⊙ AGN that is just turning on or by an exotic tidal disruption event (TDE). If the former is true, SDSS1335+0728 is one of the strongest cases of an AGNobserved in the process of activating. If the latter were found to be the case, it would correspond to the longest and faintest TDE ever observed (or another class of still unknown nuclear transient). Future observations of SDSS1335+0728 are crucial to further understand its behaviour. Key words. galaxies: active– accretion, accretion discs– galaxies: individual: SDSS J133519.91+072807.4
Synopsis presentation VDR gene polymorphism and anemia (2).pptx
Advanced Biotechnology.ppt
1. PROFESOR DR HJH FARIDA
ZURAINA MOHD YUSOF
Dean of FSG
ADVANCED MOLECULAR BIOLOGY
TECHNIQUES [OVERVIEW & TOOLS]
2. 1. Overview of Recombinant DNA technologies
a. Injection of DNA or a nucleus into a cell
b. Gene Therapy
c. “Pharm” Animals
d. Genetic Modification of Plants (e.g. GM foods)
e. Use of Prokaryotes to produce Eukaryotic gene products
3. Injection of DNA or a nucleus into Cell
Potential Applications
1. Germ line Gene Therapy—inject therapeutic gene into an egg cell (affects future generations)
2. Somatic Gene Therapy—Inject therapeutic gene into a somatic cell, culture & reinsert into an
individual
3. Cloning—inject nucleus into an enucleated egg, culture & implant into a surrogate mother.
Drawback: Inefficient means of gene transfer
4. Use of a Retrovirus
for Gene Therapy
Applications
Somatic Gene Therapy to treat
• Gaucher Disease
• SCID’s “Bubble Boy”
(Severe Combined Immune Difficiency)
5. Transgenic “Pharm” animals
Potential Applications
• Genetically modify mammals to
produce therapeutic peptide
drugs (e.g. insulin, )
• Isolate and purify drug from the
milk
• Potentially a more cost effective
method to produce
pharmaceuticals
6. Using the Ti plasmid as a vector for genetic engineering in plants
Potential Applications
Genetically modify plants to...
• produce vaccines in their fruit (e.g. polio vaccine)
• be resistant to disease and pests
• require less fertilizer, pesticides and herbicides
• have a higher nutritional value
7. “Golden” rice contrasted with ordinary rice
Transgenic Rice
• Genetically modify plants to produce beta-carotene
• Beta Carotene is converted to vitamin A in humans
• Vitamin A deficiency leads to poor vision and high susceptibility to disease
~70% of children <5 years old in SE Asia suffer from vit. A deficiency
8. Figure 20.2 An overview of how bacterial plasmids are used to clone genes
9. 2. Overview of various techniques
a. Use of Restriction Enzymes & DNA Ligase to make
recombinant DNA molecules
b. Use of Gel Electrophoresis...
• To separate restriction fragments
• For DNA fingerprinting
c. PCR (Polymerase Chain Reaction)
10. Using a restriction enzyme and DNA
ligase to make recombinant DNA
Figure 20.3
11. Gel Electrophoresis
1. A method of separating mixtures of large molecules
(such as DNA fragments or proteins) on the basis of
molecular size and charge.
2. How it’s done
• An electric current is passed through a gel containing the
mixture
• Molecules travel through the medium at a different rates
according to size and electrical charge:
Rate a size and charge
• Agarose and polyacrylamide gels are the media commonly
used for electrophoresis of proteins and nucleic acids.
15. PCR—Polymerase Chain Reaction
• A very quick, easy, automated method used to
make copies of a specific segment of DNA
• What’s needed….
1. DNA primers that “bracket” the desired sequence to be
cloned
2. Heat-resistant DNA polymerase
3. DNA nucleotides
4. Thermocycler
17. 3. Strategies used to Genetically Engineer Bacteria
See fig. 20.2. An overview of how bacterial plasmids are used to clone genes
1. Isolate the gene of interest (e.g. insulin gene)
2. Insert the gene of interest into a bacterial R-plasmid
• R-plasmids are circular DNA molecules found in some
bacteria that provide resistance to up to 10 different
antibiotics
3. Place the transgenic plasmid into bacterial cells
• Plasmid DNA reproduces each time the bacteria reproduce
4. Culture the bacteria and isolate the gene product (e.g.
insulin)
18. 3. Overview of how bacterial plasmids are used to clone genes
Figure 20.2
19. Step 1. How to Isolate the Gene of Interest
Use Reverse Transcriptase to make the gene of Interest
Method #1 (see figure on next slide)
1. Isolate mRNA for the gene product of interest (e.g. Insulin
mRNA)
2. Use Reverse Transcriptase to produce cDNA (complementary
DNA)
3. Use PCR to clone the cDNA
3. Separate the synthetic gene of interest by electrophoresis
20. Use of Reverse Transcriptase
to make complementary DNA
(cDNA) of a eukaryotic gene
21. Step 1. How to Isolate the Gene of Interest
Use Reverse Transcriptase to make the gene of Interest
Method #2
1. Determine the primary structure (i.e. the amino acid sequence)
of the protein of interest (e.g. insulin) with an automated protein
sequencer
2. Use table of codons to determine the mRNA sequence
3. Synthesize the mRNA in the lab
4. Use Reverse Transcriptase to produce cDNA and PCR to
clone the cDNA (as before)
5. Separate the synthetic gene of interest by electrophoresis
22. 1. How to Isolate the Gene of Interest
Use a labeled DNA Probe to Isolate Gene of Interest (Southern Blot Method see next slide)
1. Extract and purify DNA from cells
2. Cut DNA with restriction enzyme (e.g. Eco R1)
What’s a restriction enzyme? (fig. 20.3)
Note: Must cut outside of gene w/o too much “excess baggage”
3. Separate DNA fragments by gel electrophoresis
4. Transfer DNA from the fragile gel to a nylon sheet and heat to sep. strands (fig. 20.10)
5. Hybridize gene of interest with a radio-labeled DNA* or mRNA* probe and expose w/
film to locate gene
How do these probes work? (fig. 20.10)
6. Use PCR to clone the isolated gene of interest.
24. Steps 2 & 3. How to Insert the Gene of Interest into the R-Plasmid
See next 3 figures and animation
• Lyse bacteria with detergent to release the R-plasmid (e.g. ampicillin resistance plasmid)
• Cut the plasmid with the same restriction enzyme used to isolate the gene of interest
3. Mix plasmid with gene of interest and join the two with DNA ligase
How does this work?
4. Add the recombinant plasmid to a bacterial culture
Induce bacteria to take up plasmid (transformation)
5. Grow bacteria on agar plate containing an antibiotic (e.g. ampicillin)
6. Isolate those bacterial colonies that contain the recombinant plasmid How?
Only some of the bacteria take up a plasmid—How do you know which ones did?
Not all plasmids are recombinant plasmids—How do you find those that are?
Only some of plasmids contain the gene of interest—How do you identify these?
26. Using Plasmids to Create Recombinant DNA
1. Digest a plasmid vector with a restriction enzyme (e.g.
EcoRI) at a single site to produce two sticky ends.
2. Digest human DNA with EcoRI to produce pieces with the
same sticky ends
• Use Human DNA or cDNA copied from mRNA using reverse
transcriptase from retroviruses.
3. Mix the two samples and allow to hybridize.
• Some plasmids will hybridize with pieces of human DNA at the
EcoRI site.
4. Use DNA ligase is used to covalently link the fragments.
27. Insertion of Recombinant Plasmids into Prokaryotic Cells
1. Only some of the bacteria take
up a plasmid—How do you
know which ones did?
2. Not all plasmids are
recombinant plasmids—How
do you find those that are?
3. Only some of plasmids contain
the gene of interest—How do
you identify these?
28. Identification of cells containing plasmids
• Cells containing plasmids contain the ampicillin
resistance gene
• Grow cells on medium containing ampicillin
• How do you know which colonies contain the gene of
interest?
• Use a DNA probe (see fig. 20.5)
29. Figure 20.5
Using a DNA probe to
identify a cloned gene in a
population of bacteria
30. Step 4. Culture Bacteria and Isolate Gene Product
• Grow the recombinant bacteria in nutrient broth
and isolate/purify the gene product from the broth
• Expensive to do, therefore mammals (e.g. cows and
goats) are now being genetically modified to
produce desired gene products in their milk!!
31. Human Gene Therapy using...
a. Retroviruses
b. Adenoviruses
c. Liposomes
d. Naked DNA
32. Use of a Retrovirus
for Gene Therapy
Applications
Somatic Gene Therapy to treat
• Gaucher Disease
• SCID’s “Bubble Boy”
(Severe Combined Immune Difficiency)
33. Basic Strategies of Human Gene Therapy (1 of 2)
1. Isolate and then clone the normal allele by PCR
2. Insert normal allele into a disabled virus
• Retroviruses and adenoviruses are the most common vectors
• Retroviruses are much more efficient at forming a provirus, but have a
greater chance of mutating to cause disease
• Adenoviruses are safer, but are relatively inefficient as a vector
• Liposomes (lipid spheres) are also used as vectors
e.g. Gene therapy for Cystic Fibrosis involves using an inhaler to bring
liposomes containing the CFTR gene to the cells lining the lungs)
3. Infect host cells with recombinant virus
34. 3. Infect host cells with recombinant virus
a. Add recombinant virus directly to individual
e.g. Jesse Gelsinger—
Had Ornithine Transcarbamylase Deficiency; Causes build
up of ammonia in liver cells since they cannot convert the
ammonia (toxic) produced by amino acid metabolism to
urea (less toxic)
Died in Sept.’99 due to a severe immune response to the
genetically modified adenovirus containing the OTC gene
b. Isolate host cells from body and then add recombinant virus
(e.g. blood stem cells in gene therapy for Gaucher disease)
• Inject genetically engineered cells back into the body
Basic Strategies of Human Gene Therapy (2 of 2)