Cloning by bacterial plasmids
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Plasmid vectors are circular, self-replicating DNA molecules found in bacteria cytoplasm that can transfer genes between cells. They are useful for cloning small DNA fragments under 10kb due to their small size and selection strategies. However, plasmids are less useful for cloning large fragments over 10kb. Plasmid vectors for cloning contain an origin of replication for independent replication, selective markers for identifying cells containing the plasmid, and a multiple cloning site to insert DNA fragments.
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Vectors
Vectors are DNA molecules that can carry foreign DNA. There are two main types of vectors: cloning vectors are used to increase copies of cloned DNA fragments, while expression vectors are used to express foreign genes as proteins. Ideal vectors are autonomously replicating, contain antibiotic resistance markers, and have unique restriction sites. Common vector types include plasmids, which can clone fragments up to 10kb; bacterial artificial chromosomes for 35-300kb inserts; and yeast artificial chromosomes for 100-1000kb inserts. The choice of vector depends on the size of the DNA fragment to be cloned and the intended host organism.
Vectors in Biotechnolgy
Vectors in Biotechnolgy
- Presented By- Baisali Dora & Sonali Pati
- Guided By- Kiran Kumar Mishra,
Lecturer in Botany,
Dept. of Botany,
Vikram Dev (Auto.) College, Jeypore
Odisha, INDIA
Vector
Vectors are DNA molecules capable of self-replication that are used to introduce foreign DNA into host cells. Common vectors include plasmids, bacteriophages, cosmids, and artificial chromosomes. Vectors have features like origins of replication, selectable markers, and cloning sites that allow for replication and identification of recombinant DNA. Popular E. coli cloning vectors include the plasmids pBR322, pBR327, and pUC8, as well as the bacteriophages lambda and M13. Hybrid vectors combine characteristics of plasmids and phages, while artificial chromosomes can accommodate larger DNA fragments.
blue white selection
Gene cloning involves isolating a particular gene or DNA fragment of interest from an organism's total DNA and producing many copies of just that fragment. There are several reasons for cloning DNA, such as determining a gene's nucleotide sequence, identifying control sequences, investigating protein/enzyme function, and engineering organisms for specific purposes like insulin production. Common tools used in cloning include restriction enzymes, ligase, vectors like plasmids and bacteriophages, and host cells. DNA is cut with restriction enzymes, ligated into a vector, and introduced into host cells to replicate the exogenous DNA fragment.
Cloning vectors
This document discusses cloning vectors and their use in recombinant DNA technology. It defines a cloning vector as a DNA molecule that can accept foreign DNA and replicate within a host cell to produce multiple clones. Examples provided are plasmids, phages, cosmids, and phagemids. Key features of cloning vectors discussed include origins of replication, selectable markers, and restriction enzyme sites. Specific vectors are described in more detail, such as the E. coli plasmid pBR322, phage lambda, cosmids, and phagemids. Cloning vectors allow for the isolation of genes and determination of nucleotide sequences, as well as the investigation of protein function and identification of mutations.
Vectors part 1 | molecular biology | biotechnology
This document discusses various types of plasmid and bacteriophage vectors used for cloning DNA fragments. It describes the features and selection methods of commonly used vectors like pBR322, pUC18, pGEM3Z, lambda phage and M13 phage vectors. Plasmid vectors like pBR322 are advantageous for having small size, high copy number and antibiotic resistance markers. Lambda phage vectors can accommodate larger inserts compared to plasmids and allow easy screening of recombinant phages. M13 vectors are useful for obtaining single-stranded DNA copies for sequencing.
Vector in molecular biology
Vectors are small pieces of DNA that can accept foreign DNA and be maintained within an organism. Common vectors include plasmids, bacteriophages, and artificial chromosomes. Plasmids are circular DNA molecules that can replicate independently of the bacterial chromosome. Bacteriophages infect bacteria and have been engineered as vectors. Vectors allow for cloning of DNA fragments to study genes and their functions.
Cloning Vector
The document discusses different types of gene cloning vectors including plasmids, bacteriophages, cosmids, and phagemids. Plasmid vectors like pBR322 contain selectable markers and a multiple cloning site. Bacteriophage vectors like lambda phage infect bacteria and can carry larger DNA inserts. Cosmid vectors combine properties of plasmids and phages to clone fragments up to 50kb. All of these vector types allow cloning and replication of foreign DNA fragments in host cells.
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Vectors
Vectors are DNA molecules that can carry foreign DNA. There are two main types of vectors: cloning vectors are used to increase copies of cloned DNA fragments, while expression vectors are used to express foreign genes as proteins. Ideal vectors are autonomously replicating, contain antibiotic resistance markers, and have unique restriction sites. Common vector types include plasmids, which can clone fragments up to 10kb; bacterial artificial chromosomes for 35-300kb inserts; and yeast artificial chromosomes for 100-1000kb inserts. The choice of vector depends on the size of the DNA fragment to be cloned and the intended host organism.
Vectors in Biotechnolgy
Vectors in Biotechnolgy
- Presented By- Baisali Dora & Sonali Pati
- Guided By- Kiran Kumar Mishra,
Lecturer in Botany,
Dept. of Botany,
Vikram Dev (Auto.) College, Jeypore
Odisha, INDIA
Vector
Vectors are DNA molecules capable of self-replication that are used to introduce foreign DNA into host cells. Common vectors include plasmids, bacteriophages, cosmids, and artificial chromosomes. Vectors have features like origins of replication, selectable markers, and cloning sites that allow for replication and identification of recombinant DNA. Popular E. coli cloning vectors include the plasmids pBR322, pBR327, and pUC8, as well as the bacteriophages lambda and M13. Hybrid vectors combine characteristics of plasmids and phages, while artificial chromosomes can accommodate larger DNA fragments.
blue white selection
Gene cloning involves isolating a particular gene or DNA fragment of interest from an organism's total DNA and producing many copies of just that fragment. There are several reasons for cloning DNA, such as determining a gene's nucleotide sequence, identifying control sequences, investigating protein/enzyme function, and engineering organisms for specific purposes like insulin production. Common tools used in cloning include restriction enzymes, ligase, vectors like plasmids and bacteriophages, and host cells. DNA is cut with restriction enzymes, ligated into a vector, and introduced into host cells to replicate the exogenous DNA fragment.
Cloning vectors
This document discusses cloning vectors and their use in recombinant DNA technology. It defines a cloning vector as a DNA molecule that can accept foreign DNA and replicate within a host cell to produce multiple clones. Examples provided are plasmids, phages, cosmids, and phagemids. Key features of cloning vectors discussed include origins of replication, selectable markers, and restriction enzyme sites. Specific vectors are described in more detail, such as the E. coli plasmid pBR322, phage lambda, cosmids, and phagemids. Cloning vectors allow for the isolation of genes and determination of nucleotide sequences, as well as the investigation of protein function and identification of mutations.
Vectors part 1 | molecular biology | biotechnology
This document discusses various types of plasmid and bacteriophage vectors used for cloning DNA fragments. It describes the features and selection methods of commonly used vectors like pBR322, pUC18, pGEM3Z, lambda phage and M13 phage vectors. Plasmid vectors like pBR322 are advantageous for having small size, high copy number and antibiotic resistance markers. Lambda phage vectors can accommodate larger inserts compared to plasmids and allow easy screening of recombinant phages. M13 vectors are useful for obtaining single-stranded DNA copies for sequencing.
Vector in molecular biology
Vectors are small pieces of DNA that can accept foreign DNA and be maintained within an organism. Common vectors include plasmids, bacteriophages, and artificial chromosomes. Plasmids are circular DNA molecules that can replicate independently of the bacterial chromosome. Bacteriophages infect bacteria and have been engineered as vectors. Vectors allow for cloning of DNA fragments to study genes and their functions.
Cloning Vector
The document discusses different types of gene cloning vectors including plasmids, bacteriophages, cosmids, and phagemids. Plasmid vectors like pBR322 contain selectable markers and a multiple cloning site. Bacteriophage vectors like lambda phage infect bacteria and can carry larger DNA inserts. Cosmid vectors combine properties of plasmids and phages to clone fragments up to 50kb. All of these vector types allow cloning and replication of foreign DNA fragments in host cells.
Vectors a.k.saha
Vectors such as plasmids, lambda phage, cosmids, and yeast artificial chromosomes can be used to carry DNA fragments into host cells. Cosmids are hybrid vectors derived from plasmids that contain lambda phage cos sites, allowing them to carry 37-52 kilobase pairs of DNA. Yeast artificial chromosomes are capable of carrying large DNA fragments of up to 2 megabases but have low transformation efficiency. Plasmids are extrachromosomal DNA molecules that are important tools for cloning genes and transferring traits such as antibiotic resistance between bacteria.
Vector construction
Vectors are DNA molecules that can accept foreign DNA and be replicated within a host cell. They are required for cloning genes and transferring them to bacteria. Common vectors include plasmids, bacteriophages, cosmids, and artificial chromosomes. Expression vectors are used to produce proteins from cloned DNA, and come in prokaryotic and eukaryotic varieties. Cloning vectors replicate recombinant DNA within host cells to produce multiple clones and can accommodate different sized DNA fragments depending on the vector type.
Methods of screening
This document discusses various methods for screening recombinant clones, including:
1. Blue-white screening using X-gal to detect the presence or absence of insertions disrupting beta-galactosidase activity.
2. Insertional inactivation screening by inserting DNA fragments to disrupt antibiotic resistance or repressor genes.
3. Antibiotic sensitivity screening to detect circularized recombinant plasmids capable of conferring antibiotic resistance.
4. Auxotrophic yeast strain screening using yeast vectors containing selectable marker genes to complement host mutations.
5. Reporter gene assays using enzymes like luciferase, GFP, etc. fused to promoters to identify positive recombinant clones.
cloning vectors
cloning vectors are the kind of vehicle that is used to transfer genes from one organism to another.Different types of plasmids having different shapes are used for this purpose such as M13 plasmid, BAC, YAC, Cosmids etc
Cloning vector
Vectors are DNA molecules that can carry foreign genes into host cells. There are several types of vectors including plasmids, bacteriophages, cosmids, bacterial artificial chromosomes (BACs), and yeast artificial chromosomes (YACs). Plasmids are the most commonly used cloning vectors and can accept DNA inserts ranging from a few base pairs to 10kb. Larger DNA fragments can be cloned using vectors based on bacteriophages, cosmids, BACs, or YACs.
CLONING VECTORS SMG
Cloning vectors are DNA molecules that can carry foreign DNA fragments into recipient cells. Common types of cloning vectors include plasmids, bacteriophages, cosmids, and phagemids. Key components of vectors include an origin of replication to replicate in the host cell, selectable markers like antibiotic resistance genes to identify transformed cells, and restriction enzyme sites to insert foreign DNA. Vectors derived from bacteria like E. coli plasmid pBR322 and bacteriophage λ are widely used to clone DNA in prokaryotic cells, while plant Ti plasmids and shuttle vectors allow cloning in both prokaryotic and eukaryotic cells.
Molecular Cloning - Vectors: Types & Characteristics
Vectors are DNA molecules that can carry foreign DNA fragments into host cells. There are two main types of vectors: cloning vectors for propagating DNA inserts and expression vectors for expressing inserted DNA. Common vector types include plasmids, bacteriophages, cosmids, and artificial chromosomes. Vectors must have key characteristics like the ability to self-replicate, selectable markers, and origins of replication to successfully clone and transfer DNA.
Vectors in biotechnolgy
Vectors are DNA molecules that can carry foreign genes. This document discusses different types of vectors used in biotechnology including plasmid vectors, bacteriophage vectors, cosmid vectors, phagemid vectors, yeast vectors, animal vectors, plant vectors, and artificial chromosomes. It provides details on common vectors like pBR322 and pUC, describing their construction, uses, advantages, and disadvantages.
Vector and it's properties , types
Vectors are DNA molecules that can deliver foreign DNA into host cells. The main types of vectors are plasmids, bacteriophages, cosmids, and phagemids. Plasmids are circular, self-replicating DNA molecules that are commonly used as vectors. Key plasmid vectors include pBR322, which contains antibiotic resistance genes, and pUC18/19, which are smaller and contain an ampicillin resistance gene. Bacteriophage vectors like lambda phage and M13 phage can incorporate larger DNA fragments. Cosmids are hybrid vectors that contain phage and plasmid elements, allowing them to replicate like plasmids. Shuttle vectors contain origins of replication from different species, allowing replication in multiple host
VECTORS INTRODUCTION
This document discusses various types of vectors used for cloning and expressing plant genes, including their characteristics and uses. It describes cloning vectors, expression vectors, and integration vectors. Common vector types that are discussed in detail include plasmids, bacterial viruses (bacteriophages), cosmids, and plant viruses. The binary vector system and co-integration vector strategy for transforming plants using Agrobacterium tumefaciens are also summarized.
Vectors
Vectors can perform their functions in two main ways: transcription and expression. There are several types of vectors, including plasmids, which are small, self-replicating DNA molecules commonly used in molecular cloning. Plasmids are important tools in genetic engineering as they can be easily manipulated and transformed into bacteria to generate multiple copies of recombinant DNA. Common plasmid vectors include transcription vectors, which amplify DNA sequences without expressing proteins, and expression vectors, which are used to express foreign genes in cells.
Cloning vectors
This document discusses gene cloning and various cloning vectors. It provides details about plasmid vectors such as pBR322 and pUC18/19. pBR322 was one of the first widely used E. coli cloning vectors, containing genes for ampicillin and tetracycline resistance. It served as a model for prokaryotic transcription and replication studies. The document discusses the structure, properties, replication and uses of pBR322. It also describes the expression vector pUC18/19, which is derived from pBR322 and contains a multiple cloning site for inserting genes between the beta-lactamase gene and a fragment of the lacZ gene.
Cloning vectors
This document discusses various cloning vectors that can be used to clone DNA fragments. It describes some key vectors such as plasmids, bacteriophages, cosmids, yeast artificial chromosomes, and bacterial artificial chromosomes. Plasmids are commonly used cloning vectors that can hold inserts of up to 10kb and replicate autonomously in bacteria. Bacteriophages like lambda can accept larger inserts up to 25kb. Cosmids and YACs can hold even larger fragments, up to 40kb and over 2Mb respectively. The choice of vector depends on factors like the insert size, host system, and intended use of the cloned DNA.
Cloning vectors & gene constructs
Cloning vectors and gene constructs were submitted to Dr. Shyamalamma.S. The document discusses various topics related to molecular cloning including vectors, vector types, cloning steps, and screening procedures. Vectors are DNA molecules that can carry foreign genetic material into cells. Common vector types include plasmids, bacteriophages, cosmids, and artificial chromosomes. The cloning process involves choosing a vector and host, inserting DNA, transforming cells, and screening for positive clones.
Recombination Technology
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.
Types of vectors, Prokaryotic and Eukaryotic
vectors types
prokaryotic and eukaryotic
prokaryotic vectors= 1. ti-plasmid 2. pUC18 3. pBR322 4. lambda phage
Eukaryotic vectors= 1.BAC 2. YAC
by
Malaka Madhusudhana Reddy
Research Scholar
Dept.of :: Zoology
Yogivemana University, kadapa.
Andhrapradesh, India.
Dna libraries
1. DNA libraries contain cloned DNA fragments from an organism that can be used to isolate specific genes of interest. There are two main types: genomic libraries containing all DNA, and cDNA libraries containing expressed mRNA.
2. Bacterial transformation is a process where bacteria take up foreign DNA through horizontal gene transfer. It is used to clone and replicate plasmids in bacteria.
3. Chromosomal jumping and walking are techniques used in genome mapping and sequencing. Jumping allows bypassing repetitive sequences, while walking maps contiguous overlapping fragments to identify adjacent sequences.
ti plasmid
Ti plasmids are found in Agrobacterium tumefaciens bacteria and contain genes that allow the bacteria to transform plant cells and cause crown gall tumors. The plasmid contains virulence genes that are activated by plant signals and mediate transfer of T-DNA into the plant genome. T-DNA integration results in tumor formation and production of opines that the bacteria can utilize. Ti plasmids have been engineered as vectors for plant transformation by removing oncogenes and adding gene of interest between the border sequences, allowing transformation via Agrobacterium infection of wounded plant tissues.
Cloning plasmid
This document describes plasmids as cloning vectors. It outlines that plasmids are small, circular, self-replicating DNA molecules found in prokaryotes. Important features of plasmids for use as cloning vectors include a cloning site, origin of replication, and selectable marker like antibiotic resistance. Plasmids are useful for cloning small DNA fragments but less so for larger fragments. They provide advantages like easy selection strategies but a disadvantage is less usefulness for large DNA cloning.
Lectut btn-202-ppt-l3. gene cloning and plasmid vectors (1)
The document discusses various types of plasmid vectors and cloning vectors used for gene cloning. It describes the key properties required for a vector, including autonomous replication, small size, selectable markers, and restriction enzyme sites. Some examples of early plasmid vectors discussed are pSC101, ColE1, and pBR322. Later vectors with improved properties include the pUC series, pGEM series, and pET series. A variety of other vector types were also constructed for different applications, such as bacteriophages, cosmids, YACs, BACs, and artificial chromosomes.
DNA Cloning
This document provides an overview of DNA cloning. It begins by defining cloning as making identical copies of DNA, genes, or cells. The basic steps of DNA cloning are described, including using a source DNA, vector, restriction enzymes to cut DNA, ligation to join DNA fragments, transformation of host bacteria, and selection of recombinant clones. Common vectors like plasmids are discussed along with selection techniques like blue-white screening. The document emphasizes that the goal is to generate multiple copies of the cloned insert DNA. Examples are given of important medical and agricultural applications of cloning genes.
Plasmid technology the vector method pp
Plasmid technology uses restriction enzymes to cut viral DNA and the DNA to be inserted, fusing them together. Virulence genes are removed from viruses which are then replicated and injected, allowing the new gene to be expressed in cells and passed on through replication. Researchers have developed a viral vector delivering a gene that could cure Choroideremia with a single injection, though unpredictability of DNA insertion and using disabled viruses raise some ethical concerns. This technique aims to cure more genetic diseases in the future.
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Vectors a.k.saha
Vectors such as plasmids, lambda phage, cosmids, and yeast artificial chromosomes can be used to carry DNA fragments into host cells. Cosmids are hybrid vectors derived from plasmids that contain lambda phage cos sites, allowing them to carry 37-52 kilobase pairs of DNA. Yeast artificial chromosomes are capable of carrying large DNA fragments of up to 2 megabases but have low transformation efficiency. Plasmids are extrachromosomal DNA molecules that are important tools for cloning genes and transferring traits such as antibiotic resistance between bacteria.
Vector construction
Vectors are DNA molecules that can accept foreign DNA and be replicated within a host cell. They are required for cloning genes and transferring them to bacteria. Common vectors include plasmids, bacteriophages, cosmids, and artificial chromosomes. Expression vectors are used to produce proteins from cloned DNA, and come in prokaryotic and eukaryotic varieties. Cloning vectors replicate recombinant DNA within host cells to produce multiple clones and can accommodate different sized DNA fragments depending on the vector type.
Methods of screening
This document discusses various methods for screening recombinant clones, including:
1. Blue-white screening using X-gal to detect the presence or absence of insertions disrupting beta-galactosidase activity.
2. Insertional inactivation screening by inserting DNA fragments to disrupt antibiotic resistance or repressor genes.
3. Antibiotic sensitivity screening to detect circularized recombinant plasmids capable of conferring antibiotic resistance.
4. Auxotrophic yeast strain screening using yeast vectors containing selectable marker genes to complement host mutations.
5. Reporter gene assays using enzymes like luciferase, GFP, etc. fused to promoters to identify positive recombinant clones.
cloning vectors
cloning vectors are the kind of vehicle that is used to transfer genes from one organism to another.Different types of plasmids having different shapes are used for this purpose such as M13 plasmid, BAC, YAC, Cosmids etc
Cloning vector
Vectors are DNA molecules that can carry foreign genes into host cells. There are several types of vectors including plasmids, bacteriophages, cosmids, bacterial artificial chromosomes (BACs), and yeast artificial chromosomes (YACs). Plasmids are the most commonly used cloning vectors and can accept DNA inserts ranging from a few base pairs to 10kb. Larger DNA fragments can be cloned using vectors based on bacteriophages, cosmids, BACs, or YACs.
CLONING VECTORS SMG
Cloning vectors are DNA molecules that can carry foreign DNA fragments into recipient cells. Common types of cloning vectors include plasmids, bacteriophages, cosmids, and phagemids. Key components of vectors include an origin of replication to replicate in the host cell, selectable markers like antibiotic resistance genes to identify transformed cells, and restriction enzyme sites to insert foreign DNA. Vectors derived from bacteria like E. coli plasmid pBR322 and bacteriophage λ are widely used to clone DNA in prokaryotic cells, while plant Ti plasmids and shuttle vectors allow cloning in both prokaryotic and eukaryotic cells.
Molecular Cloning - Vectors: Types & Characteristics
Vectors are DNA molecules that can carry foreign DNA fragments into host cells. There are two main types of vectors: cloning vectors for propagating DNA inserts and expression vectors for expressing inserted DNA. Common vector types include plasmids, bacteriophages, cosmids, and artificial chromosomes. Vectors must have key characteristics like the ability to self-replicate, selectable markers, and origins of replication to successfully clone and transfer DNA.
Vectors in biotechnolgy
Vectors are DNA molecules that can carry foreign genes. This document discusses different types of vectors used in biotechnology including plasmid vectors, bacteriophage vectors, cosmid vectors, phagemid vectors, yeast vectors, animal vectors, plant vectors, and artificial chromosomes. It provides details on common vectors like pBR322 and pUC, describing their construction, uses, advantages, and disadvantages.
Vector and it's properties , types
Vectors are DNA molecules that can deliver foreign DNA into host cells. The main types of vectors are plasmids, bacteriophages, cosmids, and phagemids. Plasmids are circular, self-replicating DNA molecules that are commonly used as vectors. Key plasmid vectors include pBR322, which contains antibiotic resistance genes, and pUC18/19, which are smaller and contain an ampicillin resistance gene. Bacteriophage vectors like lambda phage and M13 phage can incorporate larger DNA fragments. Cosmids are hybrid vectors that contain phage and plasmid elements, allowing them to replicate like plasmids. Shuttle vectors contain origins of replication from different species, allowing replication in multiple host
VECTORS INTRODUCTION
This document discusses various types of vectors used for cloning and expressing plant genes, including their characteristics and uses. It describes cloning vectors, expression vectors, and integration vectors. Common vector types that are discussed in detail include plasmids, bacterial viruses (bacteriophages), cosmids, and plant viruses. The binary vector system and co-integration vector strategy for transforming plants using Agrobacterium tumefaciens are also summarized.
Vectors
Vectors can perform their functions in two main ways: transcription and expression. There are several types of vectors, including plasmids, which are small, self-replicating DNA molecules commonly used in molecular cloning. Plasmids are important tools in genetic engineering as they can be easily manipulated and transformed into bacteria to generate multiple copies of recombinant DNA. Common plasmid vectors include transcription vectors, which amplify DNA sequences without expressing proteins, and expression vectors, which are used to express foreign genes in cells.
Cloning vectors
This document discusses gene cloning and various cloning vectors. It provides details about plasmid vectors such as pBR322 and pUC18/19. pBR322 was one of the first widely used E. coli cloning vectors, containing genes for ampicillin and tetracycline resistance. It served as a model for prokaryotic transcription and replication studies. The document discusses the structure, properties, replication and uses of pBR322. It also describes the expression vector pUC18/19, which is derived from pBR322 and contains a multiple cloning site for inserting genes between the beta-lactamase gene and a fragment of the lacZ gene.
Cloning vectors
This document discusses various cloning vectors that can be used to clone DNA fragments. It describes some key vectors such as plasmids, bacteriophages, cosmids, yeast artificial chromosomes, and bacterial artificial chromosomes. Plasmids are commonly used cloning vectors that can hold inserts of up to 10kb and replicate autonomously in bacteria. Bacteriophages like lambda can accept larger inserts up to 25kb. Cosmids and YACs can hold even larger fragments, up to 40kb and over 2Mb respectively. The choice of vector depends on factors like the insert size, host system, and intended use of the cloned DNA.
Cloning vectors & gene constructs
Cloning vectors and gene constructs were submitted to Dr. Shyamalamma.S. The document discusses various topics related to molecular cloning including vectors, vector types, cloning steps, and screening procedures. Vectors are DNA molecules that can carry foreign genetic material into cells. Common vector types include plasmids, bacteriophages, cosmids, and artificial chromosomes. The cloning process involves choosing a vector and host, inserting DNA, transforming cells, and screening for positive clones.
Recombination Technology
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.
Types of vectors, Prokaryotic and Eukaryotic
vectors types
prokaryotic and eukaryotic
prokaryotic vectors= 1. ti-plasmid 2. pUC18 3. pBR322 4. lambda phage
Eukaryotic vectors= 1.BAC 2. YAC
by
Malaka Madhusudhana Reddy
Research Scholar
Dept.of :: Zoology
Yogivemana University, kadapa.
Andhrapradesh, India.
Dna libraries
1. DNA libraries contain cloned DNA fragments from an organism that can be used to isolate specific genes of interest. There are two main types: genomic libraries containing all DNA, and cDNA libraries containing expressed mRNA.
2. Bacterial transformation is a process where bacteria take up foreign DNA through horizontal gene transfer. It is used to clone and replicate plasmids in bacteria.
3. Chromosomal jumping and walking are techniques used in genome mapping and sequencing. Jumping allows bypassing repetitive sequences, while walking maps contiguous overlapping fragments to identify adjacent sequences.
ti plasmid
Ti plasmids are found in Agrobacterium tumefaciens bacteria and contain genes that allow the bacteria to transform plant cells and cause crown gall tumors. The plasmid contains virulence genes that are activated by plant signals and mediate transfer of T-DNA into the plant genome. T-DNA integration results in tumor formation and production of opines that the bacteria can utilize. Ti plasmids have been engineered as vectors for plant transformation by removing oncogenes and adding gene of interest between the border sequences, allowing transformation via Agrobacterium infection of wounded plant tissues.
Cloning plasmid
This document describes plasmids as cloning vectors. It outlines that plasmids are small, circular, self-replicating DNA molecules found in prokaryotes. Important features of plasmids for use as cloning vectors include a cloning site, origin of replication, and selectable marker like antibiotic resistance. Plasmids are useful for cloning small DNA fragments but less so for larger fragments. They provide advantages like easy selection strategies but a disadvantage is less usefulness for large DNA cloning.
Lectut btn-202-ppt-l3. gene cloning and plasmid vectors (1)
The document discusses various types of plasmid vectors and cloning vectors used for gene cloning. It describes the key properties required for a vector, including autonomous replication, small size, selectable markers, and restriction enzyme sites. Some examples of early plasmid vectors discussed are pSC101, ColE1, and pBR322. Later vectors with improved properties include the pUC series, pGEM series, and pET series. A variety of other vector types were also constructed for different applications, such as bacteriophages, cosmids, YACs, BACs, and artificial chromosomes.
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Plasmid technology the vector method pp
Plasmid technology uses restriction enzymes to cut viral DNA and the DNA to be inserted, fusing them together. Virulence genes are removed from viruses which are then replicated and injected, allowing the new gene to be expressed in cells and passed on through replication. Researchers have developed a viral vector delivering a gene that could cure Choroideremia with a single injection, though unpredictability of DNA insertion and using disabled viruses raise some ethical concerns. This technique aims to cure more genetic diseases in the future.
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Genetic engineering
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Molecular mapping
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Cloning vectors
Objectives:
After the end of the presentation we’ll know -
What is cloning vector?
Why cloning vector?
History
Features of a cloning vector
Types of cloning vector
Plasmid
Bacteriophage
Cosmid
Bacterial Artificial Chromosome (BAC)
Yeast Artificial Chromosome (BAC)
Human Artificial Chromosome (HAC)
Retroviral Vectors
What determines choice of vector?
Vector in molecular gene cloning
Cloning vector - The molecular analysis of DNA has been made possible by the cloning of DNA. The two molecules that are required for cloning are the DNA to be cloned and a cloning vector.
A cloning vector is a small piece of DNA taken from a virus, a plasmid or the cell of a higher organism, that can be stably maintained in an organism and into which a foreign DNA fragment can be inserted for cloning purposes.
Most vectors are genetically engineered.
The cloning vector is chosen according to the size and type of DNA to be cloned.
The vector therefore contains features that allow for the convenient insertion or removal of DNA fragment in or out of the vector, for example by treating the vector and the foreign DNA with a restriction enzyme and then ligating the fragments together.
After a DNA fragment has been cloned into a cloning vector, it may be further subcloned into another vector designed for more specific use.
MICROBIAL BIOTECHNOLOGY
This document discusses vector systems used in genetic engineering. It describes some key features an ideal vector should possess, including an origin of replication, selectable marker gene, unique restriction sites, small size, and expression control elements. It distinguishes between cloning vectors, meant to isolate or multiply insert DNA, and expression vectors, meant for expressing transgenes. Shuttle vectors contain origins of replication for two different hosts. The document also summarizes bacteriophage lambda's life cycle and genome organization, noting how its non-essential central region can be deleted or replaced to construct cloning vectors.
Plasmids
Plasmids are small, circular pieces of DNA found in bacteria that are separate from the main bacterial chromosome. They often contain genes for traits like drug resistance and can be transmitted between bacteria. Plasmids are used in genetic engineering and recombinant DNA techniques to transfer genes between organisms. Their discovery in the early 20th century expanded understanding of heredity, and they continue to be important tools for cloning and altering organisms through recombinant DNA. Different types of plasmids carry various traits or help with integration into chromosomes.
Cloning
Genetic engineering involves transferring genes between organisms using recombinant DNA techniques. This allows genes to be isolated, cloned, and moved within and between different species. Cloning a gene involves using restriction enzymes to cut DNA at specific sequences, and DNA ligase to join DNA fragments together. Cloned genes have many research uses such as determining gene sequences, altering phenotypes, and obtaining protein products of genes.
Dna cloning intro
DNA cloning allows for the replication and amplification of specific DNA fragments. It involves inserting DNA fragments into vector molecules, such as plasmids, which are then introduced into host cells. The vector carries the DNA fragment into the host cell and acts as a replicating molecule. Restriction enzymes and ligases are used to cut out the DNA fragment and insert it into the vector. The cells are then grown on selective media so that cells containing the vector and insert are selected for. This allows unlimited copies of the DNA fragment of interest to be produced for study and manipulation.
Molecular cloning vectors
The molecular cloning vectors used in genetic engineering-plasmids,phage vectors,Hbrid vectors and artificial chromosomes
Crispr/cas9 101
This document provides an overview of CRISPR/Cas9 genome editing. It discusses the history and limitations of prior genome engineering techniques like recombinant DNA and zinc finger nucleases. It then explains how CRISPR/Cas9 works as a RNA-guided DNA endonuclease and how this allows it to efficiently and specifically edit genomes. The document outlines several applications of CRISPR/Cas9 like generating knockout animals and cell lines. It also notes some concerns about using the technique for human genome editing.
Gene cloning
Gene cloning involves making many identical copies of a gene. It differs from cloning an entire organism. Gene cloning is done to isolate and study particular genes, identify control sequences and mutations, and engineer organisms for specific purposes like insulin production. It is accomplished by extracting DNA from an organism, cutting it into pieces with restriction enzymes, and inserting the gene of interest into bacterial plasmids which are then taken up by bacteria. This allows the gene to be amplified as the bacteria containing the recombinant plasmid multiply. PCR is another method that amplifies DNA fragments in vitro using heat-stable DNA polymerase and repeated heating and cooling cycles.
Bacterial plasmids
plasmids
history
properties
classification
ColE1 plasmid
characteristics
replication
regulation
application
Sv 40 plasmid
history
properties
applications
PMB 9 Plasmid
construction
application
Bacteriophage vectors
Bacteriophage vectors
Bacteriophage
WHY BACTERIOPHAGE AS A VECTOR?
M13 phage
Genome of m13 phage
Life cycle and dna replication of m13
CONSTRUCTION M13 AS PHAGE VECTOR
M13 MP 2 vector
M13MP7 VECTOR
Selection of recombinants
Lambda replacement vectors
LAMBDA EMBL 4 VECTOR
P1 PHAGE
GENOME OF P1 PHAGE
P1 PHAGE AS VECTOR
P1 phage vector system
Vectors
This document discusses various types of vectors used for cloning, including bacteriophage vectors, plasmid vectors, cosmid vectors, bacterial artificial chromosomes (BACs), yeast artificial chromosomes (YACs), and shuttle vectors. Key points include:
- Bacteriophage derivatives like lambda phage are suitable for cloning large eukaryotic DNA due to abilities like packaging millions of clones and size selection of DNA.
- Phage-based vectors can be insertional, containing a single cloning site, or replacement vectors with two cloning sites allowing DNA substitution.
- Cosmids are hybrid phage-plasmid vectors that can package DNA up to 48 kb into phage particles.
- Y
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Cloning vectors are small DNA molecules used to replicate, amplify and express inserted DNA fragments. There are several types of cloning vectors including plasmids, bacteriophages, cosmids, and artificial chromosomes. Plasmids are the most commonly used cloning vectors as they can replicate autonomously in bacterial cells, contain selectable markers, and accept DNA insert sizes up to 10kb. Bacteriophages such as lambda can accept larger inserts up to 20kb but have a narrow host range. Cosmids combine properties of plasmids and phages to accept inserts up to 50kb.
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The document discusses different types of cloning vectors. It describes cloning vectors as DNA fragments capable of self-replication that can transport foreign genetic material into host cells. The main types discussed are plasmids, bacteriophages, cosmids, BACs, YACs, and retroviral vectors. Each type has distinct features like replication origin and cloning capacity that determine their suitability for different applications. Plasmids were among the earliest vectors and can clone inserts up to 10kb, while BACs and YACs can accommodate much larger DNA fragments. The key factors in choosing a vector are the size of the DNA insert and cloning efficiency required.
Recombinant DNA technology
Vectors are used to carry DNA fragments into host cells for replication. Plasmids are commonly used cloning vectors that are extrachromosomal and autonomously replicating in bacteria. Properties of good vectors include an origin of replication, antibiotic resistance marker, and unique restriction sites. Different vector types include plasmids, bacteriophages, cosmids, BACs, YACs, and mini chromosomes, each with advantages for cloning varying size DNA fragments. Recombinant DNA technology uses restriction enzymes to cut DNA, ligase to join fragments, and vectors to replicate, express, and select for cloned genes.
Cloning vectors
It is the basics of vector cloning which necessary for every and each student who is intrested in biotechnology. It is only starting, if you want to more than this then please comment on it.
cloning vectors.ppt
Cloning vectors are small DNA molecules that can accept foreign DNA inserts and replicate within a host cell. They contain features like an origin of replication, antibiotic resistance genes, and restriction enzyme cleavage sites. Common vector types include plasmids, bacteriophages, cosmids, and artificial chromosomes. The choice of vector depends on factors like the size of the DNA insert and the intended use. Vectors allow amplification and manipulation of cloned DNA fragments.
Gene cloning
GENE CLONING
PLASMIDS
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cloning vectors,
cosmid properties,
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quality of good vectors,
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Comparison Between Different Types Of Vectors
This document compares different types of cloning vectors, including plasmid vectors like pBR322 and pUC19, bacteriophage lambda cloning vectors, cosmid vectors, and shuttle vectors. Plasmid vectors are small, circular DNA molecules that can carry foreign DNA fragments of up to 10kb. Lambda bacteriophage vectors can accept larger inserts up to 15kb. Cosmid vectors can carry the largest foreign DNA fragments, up to 40kb, and shuttle vectors allow propagation in two different host species. Each vector type has advantages like insert size capacity and disadvantages like difficulty of manipulation.
Cloning Vectors (plasmid).pdf
Plasmids are circular, self-replicating DNA molecules that are commonly used as cloning vectors. Key properties of plasmids that make them useful for cloning include their ability to replicate independently of the bacterial chromosome, carry genes for antibiotic resistance or other selectable markers, and accept foreign DNA at specific restriction sites. Common plasmids used for cloning are pBR322, which has features like a high copy number and blue-white screening, and pUC plasmids, which have even higher copy numbers. Plasmids can shuttle DNA between bacterial and yeast cells, and RNA production plasmids contain promoters to transcribe cloned DNA into RNA for applications like Northern blotting.
cloning vectors.pptx Biotechnology Notes
A cloning vector is a small piece of DNA, such as a plasmid, virus, or artificial chromosome, that can accept foreign DNA and be used to clone that DNA and replicate it in a host cell. This document discusses the history and features of common cloning vectors like plasmids, bacteriophages, cosmids, and artificial chromosomes. It explains how vectors are chosen based on factors like insert size and used in molecular cloning by digesting DNA with restriction enzymes, ligating into the vector, transforming into host cells, and selecting for recombinant clones.
lecture-2_4.pptx
Cloning vectors such as plasmids, bacteriophages, bacterial artificial chromosomes (BACs), yeast artificial chromosomes (YACs), and mammalian artificial chromosomes (MACs) are used to clone DNA fragments of varying sizes. Common features of vectors include being self-replicating inside host cells, containing unique restriction sites and selectable marker genes. Plasmids are small, circular DNA molecules that can clone up to 10kb fragments into bacteria. Bacteriophages infect bacteria and can deliver larger fragments up to 20kb. BACs stably clone fragments up to 300kb in bacteria. YACs clone very large fragments from 100kb to 3Mb in yeast and allow eukaryotic expression.
1. Study of cloning vectors, restriction endonucleases and DNA ligase.pptx
Vectors are DNA molecules that transport foreign DNA into host cells. Common vectors include plasmids, bacteriophages, cosmids, and artificial chromosomes. Cloning vectors are used to replicate and express foreign DNA in host organisms. Restriction enzymes cut DNA at specific recognition sites, while DNA ligase joins cut DNA fragments together. These tools enable cloning of DNA fragments into vectors for amplification and study.
Plasmid
This document provides an overview of plasmids. It defines plasmids as small, circular, extrachromosomal DNA molecules that can replicate independently in bacteria. Plasmids contain genes that provide benefits to bacteria like antibiotic resistance. They are transferred between bacteria through processes like transformation, transduction, and conjugation. Plasmids are classified based on their functions and are important tools in biotechnology as they allow cloning, protein production, and other applications.
LECTURE 5_Principle of Cloning.pdf
Cloning involves creating identical copies of DNA fragments. Molecular cloning uses restriction enzymes to cut DNA into fragments which are then joined to plasmid vectors via ligation. The recombinant plasmids are introduced into host bacteria via transformation. This allows the DNA fragment to be amplified in large quantities for further study and manipulation. Common vectors used in cloning include plasmids, bacteriophages, cosmids, and YACs which can accommodate varying sizes of DNA inserts.
DNA_cloning_principles and procedures.ppt
This document discusses principles of DNA cloning including DNA cloning techniques, vectors, and cloning strategies. DNA cloning allows for amplification of specific DNA fragments and involves inserting DNA fragments into vectors, such as plasmids, for propagation in host cells. Key steps in plasmid cloning include restriction enzyme digestion of the DNA fragment and vector, ligation of the fragment into the vector, transformation of host cells, and selection of cells containing the recombinant DNA. Common vectors include plasmids, bacteriophages, cosmids, and yeast artificial chromosomes which can accommodate varying sizes of DNA inserts.
Principles of cloning DNA introduction
This document discusses principles and strategies for cloning DNA fragments. It describes how DNA cloning allows copies of specific DNA sequences to be produced in unlimited amounts. The key steps involve using restriction enzymes to cut DNA fragments and vectors, ligating the fragment into the vector, transforming host cells, and selecting clones. Common vectors discussed are plasmids, bacteriophages, cosmids, and artificial chromosomes. The document outlines the plasmid cloning strategy involving restriction digestion, ligation, transformation, and blue/white screening to select recombinant clones.
PLANT TRANSFORMATIVE VECTORS.pptx
Plant transformation vectors can be classified into cloning vectors, expression vectors, and integration vectors. Cloning vectors are small DNA molecules used to insert, store, and manipulate foreign DNA. Common cloning vectors include plasmids, bacteriophages, cosmids, and BACs/YACs. Expression vectors allow foreign DNA to be inserted and expressed in host cells. The Ti plasmid is often used as a plant expression vector due to genes that mediate DNA transfer to plant cells. Vector choice depends on desired DNA insert size, host system, and purpose of cloning/expression.
PLANT TRANSFORMATIVE VECTORS.pptx
Plant transformation, vectors, vectors classification, cloning vectors, types of cloning vectors, plasmid vectors, classification of plasmid vectors, pBR322, advantages and disadvantages of vectors, Bacteriophage vectors, phage Lambda, M13 vectors, cosmids, bacterial artificial chromosome, yeast artificial chromosome, shuttle vectors, expression vectors, Ti plasmid, its advantages and disadvantage, conclusion
Cloning vectors
This document discusses cloning vectors. It begins by defining a cloning vector as a small piece of DNA that can stably maintain and replicate an inserted foreign DNA fragment for cloning purposes. It then lists some key properties of cloning vectors, including autonomous replication, restriction enzyme sites, and selectable marker genes. The main types of cloning vectors discussed are plasmids, bacteriophages, cosmids, BACs, YACs, HACs, and shuttle/expression vectors. One specific example, the pBR322 plasmid vector, is then described in more detail.
Lecture_3-Vectors.pdf
The document discusses cloning vectors. It describes what a cloning vector is, including that it is a small piece of DNA that can stably maintain foreign DNA for cloning purposes. Common types of cloning vectors are described in detail, including plasmids, bacteriophages, cosmids, yeast artificial chromosomes, bacterial artificial chromosomes, and plant virus vectors. Key features of cloning vectors like origins of replication, antibiotic resistance genes, and cloning sites are also summarized.
toolsusedingeneticengineeringnpk-170305183111.pdf
This document summarizes various tools used in genetic engineering. It describes restriction enzymes like EcoRI and HinDIII that cut DNA at specific sequences. It also discusses DNA ligases that join DNA fragments and alkaline phosphatase which removes phosphates to prevent unwanted ligation. Common vectors like plasmids, bacteriophages, cosmids, BACs and YACs are outlined which allow insertion of exogenous DNA for storage and manipulation. Plasmids are described as circular DNA that can replicate autonomously in bacteria while bacteriophages like lambda can accept short DNA fragments. BACs and YACs can hold larger fragments up to 300kb and 500kb respectively.
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Cloning by bacterial plasmids
- 1. Cloningcloning by bacterial plasmids PRESENTED BY BUMBIM NO.20 FAH NO.32
- 2. Plasmid vectors are double-stranded, circular, self-replicating, extra-chromosomal DNA molecules. Advantages: • Small, easy to handle • Straightforward selection strategies • Useful for cloning small DNA fragments (< 10kbp) Disadvantages: • Less useful for cloning large DNA fragments (> 10kbp) Plasmid vectors
- 3. Plasmids are circular DNA molecules present in the cytoplasm of the bacteria Capable of autonomous replication Can transfer genes from one cell to other Act as vectors in genetic engineering Can also present in Yeasts Plasmid vectors
- 4. may encode genetic information for properties 1 Resitance to Antibiotics 2 Bacteriocins production 3 Enterotoxin production 4 Enhanced pathogen city 5 Reduced Sensitivity to mutagens 6 Degrade complex organic molecules Plasmid vectors
- 5. Plasmid vector for cloning 1.Contains an origin of replication, allowing for replication independent of host’s genome. 2.Contains Selective markers: Selection of cells containing a plasmid twin antibiotic resistance blue-white screening 3.Contains a multiple cloning site (MCS) 4.Easy to be isolated from the host cell.