Genetic engineering
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This document summarizes recombinant DNA (rDNA) technology and genetic engineering. It discusses how restriction enzymes and DNA ligases are used to cut and paste DNA from different sources to create recombinant DNA molecules. It also describes genetically modified organisms (GMOs) and provides examples of transgenic organisms. Vectors such as plasmids are used to carry and replicate foreign DNA fragments in bacteria. Selection techniques like antibiotic resistance allow identification of transformed bacteria.
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Applications of Recombinant DNA Technology
Recombinant DNA technology allows for the creation of novel gene combinations not found in nature. Genes can be inserted into host cells and replicated. This technology is used in research, medicine, agriculture, and industry. In medicine, it enables production of insulin, growth factors, vaccines, and gene therapy. It is also used for genetic mapping, disease diagnosis, forensics, and prenatal testing. In agriculture, it improves crop yields and develops pest/drought resistance. Industry utilizes it to produce enzymes, additives, sugars, and chemicals.
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Lysozyme is an enzyme that catalyzes the breakdown of bacterial cell walls. It was first discovered in 1922 by Alexander Fleming. The enzyme hydrolyzes beta linkages between N-acetylglucosamine and N-acetylmuramic acid in peptidoglycan. Lysozyme has antibacterial properties and is found naturally in human tears, saliva, and egg whites. It has important applications in protein purification and nucleic acid preparation. The enzyme has a compact globular structure and contains four disulfide bonds that provide stability. Its active site contains the residues glutamic acid 35 and aspartic acid 52, which work together to catalyze the hydrolysis reaction.
Molecular chaperones
This document provides an overview of molecular chaperones. It defines chaperones as proteins that assist in protein folding but are not part of the final structure. Chaperones exist in all cellular compartments and help prevent protein misfolding and aggregation. They are classified as intra-molecular or inter-molecular. The best characterized system is the GroEL/GroES complex which uses ATP to encapsulate proteins and facilitate folding. Chaperones have implications for developing treatments for diseases like cancers and lysosomal storage disorders.
Biochemistry - Ch4 protein structure , and function
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G protein coupled receptors (Type II Hormone Signaling)
G Protein–Coupled Receptors (GPCRs) are integral membrane proteins that are activated by extracellular signaling molecules and activate intracellular secondary messenger pathways. They have seven transmembrane domains and activate heterotrimeric G proteins upon ligand binding. The G protein then activates downstream effector enzymes like adenylyl cyclase, which generates secondary messengers like cAMP. These messengers go on to activate pathways that ultimately alter cell function. The beta-adrenergic receptor pathway is a key example, with epinephrine binding and cAMP production leading to protein kinase A activation. PKA then phosphorylates target proteins to produce effects like increased heart rate. GPCRs are major drug targets, and their deregulation
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Signaling molecules
Cell signaling involves the use of signaling molecules to transmit information between cells. These molecules can be classified as extracellular signals, like peptides, lipids, gases, and small hydrophilic molecules, or intracellular second messengers like cAMP and calcium. Extracellular signals bind to cell surface receptors and trigger intracellular pathways that regulate cell function and development. Signaling can occur through endocrine, paracrine, or autocrine pathways depending on the distance over which the signal acts. Important examples of signaling molecules discussed include peptide hormones, steroid hormones, prostaglandins, and nitric oxide. Intracellular signaling molecules like G proteins and protein kinases transmit and amplify extracellular signals within cells through the use of feedback loops and molecular switches. Breakdowns
Recommended
Applications of Recombinant DNA Technology
Recombinant DNA technology allows for the creation of novel gene combinations not found in nature. Genes can be inserted into host cells and replicated. This technology is used in research, medicine, agriculture, and industry. In medicine, it enables production of insulin, growth factors, vaccines, and gene therapy. It is also used for genetic mapping, disease diagnosis, forensics, and prenatal testing. In agriculture, it improves crop yields and develops pest/drought resistance. Industry utilizes it to produce enzymes, additives, sugars, and chemicals.
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Lysozyme
Lysozyme is an enzyme that catalyzes the breakdown of bacterial cell walls. It was first discovered in 1922 by Alexander Fleming. The enzyme hydrolyzes beta linkages between N-acetylglucosamine and N-acetylmuramic acid in peptidoglycan. Lysozyme has antibacterial properties and is found naturally in human tears, saliva, and egg whites. It has important applications in protein purification and nucleic acid preparation. The enzyme has a compact globular structure and contains four disulfide bonds that provide stability. Its active site contains the residues glutamic acid 35 and aspartic acid 52, which work together to catalyze the hydrolysis reaction.
Molecular chaperones
This document provides an overview of molecular chaperones. It defines chaperones as proteins that assist in protein folding but are not part of the final structure. Chaperones exist in all cellular compartments and help prevent protein misfolding and aggregation. They are classified as intra-molecular or inter-molecular. The best characterized system is the GroEL/GroES complex which uses ATP to encapsulate proteins and facilitate folding. Chaperones have implications for developing treatments for diseases like cancers and lysosomal storage disorders.
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This document discusses the structure of proteins at multiple levels. It explains that a protein's amino acid sequence determines its 3D structure, and its structure dictates its function. Noncovalent interactions like hydrogen bonds, hydrophobic interactions, and electrostatic interactions are important forces that stabilize a protein's native structure. The peptide bond is rigid and planar, limiting the possible conformations of the polypeptide backbone. Common secondary structures like alpha helices and beta sheets form due to favorable hydrogen bonding patterns between peptide bonds.
G protein coupled receptors (Type II Hormone Signaling)
G Protein–Coupled Receptors (GPCRs) are integral membrane proteins that are activated by extracellular signaling molecules and activate intracellular secondary messenger pathways. They have seven transmembrane domains and activate heterotrimeric G proteins upon ligand binding. The G protein then activates downstream effector enzymes like adenylyl cyclase, which generates secondary messengers like cAMP. These messengers go on to activate pathways that ultimately alter cell function. The beta-adrenergic receptor pathway is a key example, with epinephrine binding and cAMP production leading to protein kinase A activation. PKA then phosphorylates target proteins to produce effects like increased heart rate. GPCRs are major drug targets, and their deregulation
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Signaling molecules
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Acid –Base Properties of Amino Acids
- Amino acids exist in two forms, L and D, depending on their stereoisomer configuration. Only L-amino acids are used in protein synthesis.
- Amino acids are amphoteric, meaning they can act as acids or bases depending on the pH. At low pH they are protonated and at high pH they are deprotonated.
- In solution, amino acids exist as zwitterions - molecules with both positive and negative charges that cancel out to have no overall charge. The isoelectric point is the pH at which the zwitterion form dominates.
membrane protein, synthesis by
Introduction
Protein synthesis
Synthesis of secretory proteins on membrane-bound ribosomes
Processing of newly synthesized proteins in the ER
Synthesis of integral membrane protein on membrane bound ribosomes
Maintenance of membrane asymmetry
Conclusion
Reference
Protein kinases
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1. Protein moonlighting refers to the phenomenon where a protein can perform more than one function. Many proteins have been found to "moonlight" and take on secondary functions beyond their primary role.
2. Examples of moonlighting mechanisms include differential localization, expression, oligomerization, complex formation, and ligand/substrate concentrations. Moonlighting proteins are often highly conserved and can help expand an organism's functional capabilities without increasing genome size.
3. Identifying moonlighting proteins can provide insights into complex disease phenotypes and bacterial virulence properties. Further research using structural analysis, interactomics, and machine learning approaches may help examine these multifunctional proteins.
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Protein sturucture
This document discusses protein structure and function. It begins by defining proteins and their essential roles in the human body. It then describes the general characteristics of proteins, such as their chemical composition and functions. The document classifies proteins based on their composition, axial ratio, and biological functions. It explains the different levels of protein structure from primary to quaternary, focusing on secondary structure elements like alpha helices and beta sheets. The roles of hydrogen bonding, disulfide bridges, and ionic bonds in tertiary and quaternary structure are also described.
Plasmids
Plasmids are small, circular, self-replicating DNA molecules found in bacterial cells separate from the chromosomal DNA. They can replicate independently and be transferred between bacteria. Plasmids are classified based on their ability to transfer (conjugative or non-conjugative) and their functions like F and F' plasmids involved in bacterial conjugation, R plasmids conferring antibiotic resistance, and virulence plasmids making bacteria pathogenic. Plasmids replicate through rolling circle replication or by displacing one strand of DNA to form a displacement loop. They are important for medical uses like producing antibiotics, agricultural uses like engineering herbicide/pest resistance in crops, and industrial applications including producing enzymes and recombinant proteins.
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Slideshare second messengers aj
1. Second messengers are small intracellular molecules that transmit signals within cells after extracellular signaling molecules (hormones or neurotransmitters) bind to cell surface receptors.
2. There are three main types of second messenger systems: cyclic AMP (cAMP), cyclic GMP (cGMP), and inositol trisphosphate (IP3)/diacylglycerol (DAG). These systems activate protein kinases or trigger the release of calcium ions to produce a physiological response.
3. Second messengers amplify and diversify extracellular signals, allowing for precise regulation of multiple cellular processes. Their roles are important for understanding cell signaling, disease mechanisms, and potential drug targets.
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Types of transport include passive transport which does not require energy and active transport which uses protein pumps and channels to move substances across membranes. There are several types of active transport including bulk transport, endocytosis, and exocytosis. Endocytosis brings material into the cell through phagocytosis, pinocytosis, or receptor-mediated endocytosis while exocytosis expels material from the cell. Phagocytosis engulfs solid particles, pinocytosis brings in extracellular fluid through membrane invagination, and receptor-mediated endocytosis specifically uptakes substances bound to cell surface receptors. Both endocytosis and exocytosis use vesicle formation and membrane fusion but transport materials in opposite directions with endocytosis importing and exocytosis exporting.
Cell signaling pathways
Cell signaling pathways allow cells to communicate with each other. Signaling occurs through the release of signal molecules called ligands that bind to receptors on other cells. There are different types of cell signaling including paracrine signaling between nearby cells, endocrine signaling over long distances via hormones in the bloodstream, and autocrine signaling where a cell secretes and responds to its own signals. Cell signaling pathways play an important role in coordinating complex processes in multicellular organisms.
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This document provides an overview of recombinant DNA technology. It discusses the basic principles, which involve generating DNA fragments, inserting a selected fragment into a cloning vector, introducing the vector into host cells, and multiplying the recombinant molecules. Key steps include using restriction enzymes to cut DNA at specific sites, ligases to join fragments, and various vectors like plasmids and bacteria to clone the DNA. The document also outlines several applications of rDNA technology, such as producing proteins, diagnosing diseases, and developing genetically engineered plants.
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Acid –Base Properties of Amino Acids
- Amino acids exist in two forms, L and D, depending on their stereoisomer configuration. Only L-amino acids are used in protein synthesis.
- Amino acids are amphoteric, meaning they can act as acids or bases depending on the pH. At low pH they are protonated and at high pH they are deprotonated.
- In solution, amino acids exist as zwitterions - molecules with both positive and negative charges that cancel out to have no overall charge. The isoelectric point is the pH at which the zwitterion form dominates.
membrane protein, synthesis by
Introduction
Protein synthesis
Synthesis of secretory proteins on membrane-bound ribosomes
Processing of newly synthesized proteins in the ER
Synthesis of integral membrane protein on membrane bound ribosomes
Maintenance of membrane asymmetry
Conclusion
Reference
Protein kinases
Protein kinases are enzymes that phosphorylate other proteins and cause functional changes in target proteins. There are over 500 types of protein kinases in genomes that can modify about 30% of proteins. Protein kinase A, C, and G are three important protein kinases that are regulated by different mechanisms and have various functions in different cell types. Protein kinases play critical roles in many cellular processes and their dysregulation can lead to diseases like cancer.
Extracellular matrix
The document provides an overview of the two main classes of extracellular matrix (ECM) molecules - glycosaminoglycan polysaccharide chains and fibrous proteins. Glycosaminoglycans form hydrated gels that allow for nutrient diffusion and withstand compressive forces. Fibrous proteins organize and strengthen the ECM by forming networks with other ECM components. The ECM is organized into connective tissue and basal lamina and interacts with cells through integrins.
4 signal ligand gated ion channels.
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Expression systems
Commercial production of gene products requires high levels of gene expression. Several factors can be manipulated to increase protein production, including the vector, host chromosome location, gene dosage, transcription elements like promoters and terminators, translation elements like ribosome binding sites, and codon optimization. Final localization of the protein, such as secretion extracellularly, and preventing degradation through fusion proteins or other means can also improve yields. However, expression in E. coli has limitations like the inability to perform post-translational modifications and potential endotoxin contamination.
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Calcium signalling occurs due to changes in cytosolic calcium concentration and activates protein kinase C. Protein kinase C is activated when phospholipase C breaks down phospholipids like phosphatidylinositol, producing diacylglycerol which remains in the cell membrane and recruits protein kinase C to the membrane. This allows calcium-calmodulin complexes to activate protein kinase C, leading to phosphorylation of transcription factors and regulation of processes like cell growth and metabolism. Phospholipase C also produces IP3 which triggers release of calcium from intracellular stores, further increasing cytosolic calcium levels.Protein moonlighting
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3. Identifying moonlighting proteins can provide insights into complex disease phenotypes and bacterial virulence properties. Further research using structural analysis, interactomics, and machine learning approaches may help examine these multifunctional proteins.
History of enzymes.
This document provides a history of enzymes, beginning with early references to enzymatic processes in ancient texts. It describes key discoveries such as Kirchoff's observation of starch hydrolysis, Pasteur's conclusion that fermentation requires living yeast cells, and Buchner's demonstration of cell-free fermentation. Major milestones include the first isolation of an enzyme (diastase) in 1833, Sumner's purification and crystallization of urease in 1926, and the determination of enzyme structures using X-ray crystallography beginning in 1965. The document also notes the discovery of ribozymes in the 1980s and restriction enzymes in the 1960s-1970s.
Protein sturucture
This document discusses protein structure and function. It begins by defining proteins and their essential roles in the human body. It then describes the general characteristics of proteins, such as their chemical composition and functions. The document classifies proteins based on their composition, axial ratio, and biological functions. It explains the different levels of protein structure from primary to quaternary, focusing on secondary structure elements like alpha helices and beta sheets. The roles of hydrogen bonding, disulfide bridges, and ionic bonds in tertiary and quaternary structure are also described.
Plasmids
Plasmids are small, circular, self-replicating DNA molecules found in bacterial cells separate from the chromosomal DNA. They can replicate independently and be transferred between bacteria. Plasmids are classified based on their ability to transfer (conjugative or non-conjugative) and their functions like F and F' plasmids involved in bacterial conjugation, R plasmids conferring antibiotic resistance, and virulence plasmids making bacteria pathogenic. Plasmids replicate through rolling circle replication or by displacing one strand of DNA to form a displacement loop. They are important for medical uses like producing antibiotics, agricultural uses like engineering herbicide/pest resistance in crops, and industrial applications including producing enzymes and recombinant proteins.
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Lineweaver - Burk plot.pptx
The document discusses Lineweaver-Burk plots, which are double reciprocal plots used to transform the Michaelis-Menten equation for enzyme kinetics into a linear form. This allows for a more accurate determination of the maximum velocity (Vmax) and Michaelis constant (Km) of an enzymatic reaction. The document provides examples of how to construct Lineweaver-Burk plots and extract Km and Vmax values from them. It also discusses how the plots can be used to determine the type of inhibition occurring in the presence of an inhibitor.
cytosceleton
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1. Second messengers are small intracellular molecules that transmit signals within cells after extracellular signaling molecules (hormones or neurotransmitters) bind to cell surface receptors.
2. There are three main types of second messenger systems: cyclic AMP (cAMP), cyclic GMP (cGMP), and inositol trisphosphate (IP3)/diacylglycerol (DAG). These systems activate protein kinases or trigger the release of calcium ions to produce a physiological response.
3. Second messengers amplify and diversify extracellular signals, allowing for precise regulation of multiple cellular processes. Their roles are important for understanding cell signaling, disease mechanisms, and potential drug targets.
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This document summarizes enzyme engineering. Enzyme engineering is the process of designing enzymes by changing the amino acid sequence through recombinant DNA technology. This allows modifying enzyme properties for industrial and other applications, such as improved kinetics, thermostability or pH suitability. There are two main methods - rational design which uses structural knowledge to make specific changes, and directed evolution which mimics natural evolution in vitro through random mutagenesis and selection to evolve enzymes with desired properties.
Exocytosis and Endocytosis {Best one}
Types of transport include passive transport which does not require energy and active transport which uses protein pumps and channels to move substances across membranes. There are several types of active transport including bulk transport, endocytosis, and exocytosis. Endocytosis brings material into the cell through phagocytosis, pinocytosis, or receptor-mediated endocytosis while exocytosis expels material from the cell. Phagocytosis engulfs solid particles, pinocytosis brings in extracellular fluid through membrane invagination, and receptor-mediated endocytosis specifically uptakes substances bound to cell surface receptors. Both endocytosis and exocytosis use vesicle formation and membrane fusion but transport materials in opposite directions with endocytosis importing and exocytosis exporting.
Cell signaling pathways
Cell signaling pathways allow cells to communicate with each other. Signaling occurs through the release of signal molecules called ligands that bind to receptors on other cells. There are different types of cell signaling including paracrine signaling between nearby cells, endocrine signaling over long distances via hormones in the bloodstream, and autocrine signaling where a cell secretes and responds to its own signals. Cell signaling pathways play an important role in coordinating complex processes in multicellular organisms.
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rDNA technology
Recombinant DNA technology involves generating DNA fragments from different sources in vitro, cutting and joining the fragments to vector DNA molecules, and introducing the vectors carrying the foreign DNA into host cells where they can replicate and selecting clones that have acquired the recombinant DNA.
Recombinant DNA or rDNA
Recombinant DNA refers to DNA molecules formed by combining DNA from different sources. It is produced artificially by joining DNA from different organisms. The pioneers who developed recombinant DNA technology were Paul Berg, Herbert Boyer, and Stanley Cohen in the 1970s. The process involves using restriction enzymes to cut DNA fragments, which are then joined to vector DNA and inserted into host cells where they can be replicated. This allows scientists to modify genes and study their functions.
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1. Recombinant DNA technology involves manipulating DNA from different species and combining them to form new recombinant DNA molecules.
2. Key steps include using restriction enzymes to cut DNA at specific sites, and DNA ligase to join DNA fragments together into vectors like plasmids.
3. The recombinant DNA can then be replicated in host cells like bacteria to produce multiple copies for analysis.
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Genetic engineering involves transferring DNA between organisms. It uses recombinant DNA techniques where the gene of interest is isolated and inserted into a vector like a plasmid or virus, which is then used to introduce the gene into a host cell. This allows the production of useful proteins like insulin through genetically modified bacteria. While genetic engineering has benefits like producing important medicines, there are also potential health and environmental risks to consider.
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Joshua lederberg ppt
Joshua Lederberg was born in 1925 in New Jersey and showed a strong interest in science from a young age, influenced by books like The Microbe Hunters. He graduated early from the specialized science high school Stuyvesant and continued experiments at the American Institute Science Laboratory. He then attended Columbia University, where he studied under mentor Francis Ryan and became interested in using chemical analysis to study life through the mold Neurospora. Lederberg went on to make pioneering contributions to the new fields of bacterial genetics and molecular biology.
recombinant DNA with subtopics
Recombinant DNA (rDNA) refers to DNA molecules formed by combining genetic material from multiple sources. Basic techniques for creating rDNA include using restriction enzymes to cut DNA at specific sites, vectors to transport DNA into host cells, and DNA ligase to join DNA fragments. Important applications of rDNA technology include producing insulin, vaccines, and proteins as well as creating pest-resistant plants. Site-directed mutagenesis allows researchers to precisely introduce mutations at specific locations in DNA, while random mutagenesis introduces random mutations throughout DNA.
Biotech
In 1973, Herbert Boyer and Stanley Cohen reported the first successful construction of organisms that combined genetic information from different species. They inserted genes from the African clawed frog Xenopus laevis into bacteria, demonstrating that these genes were active across generations of E. coli. This achievement represented a major advance in genetic engineering and showed that genes and gene products could now be rapidly produced in large quantities using bacteria. Their work built upon earlier research developing techniques for inserting viral DNA into bacterial DNA and marked a watershed moment, launching the field of genetic engineering and biotechnology.
10 mutation
Mutation is a change in DNA base sequence that can alter protein function. Spontaneous mutation rates vary from 10-6 to 10-9 per gene. There are two main types of mutations: substitutions of single bases and insertions/deletions of multiple bases. Substitutions can be silent, missense, or nonsense. Insertions and deletions cause frameshift mutations. Mutagens like chemicals, radiation, and viruses increase mutation rates. Mutants are detected by direct selection on media that allows growth of mutants but not parents, or by enrichment with penicillin or replicate plating.
BACTERIAL RECOMBINATION, PLASMIDS AND EPISOMES
Bacterial genetic recombination can occur through transformation, transduction, or conjugation. Transformation involves DNA uptake from dead bacteria. Transduction involves DNA transfer by bacteriophages. Conjugation involves DNA transfer through cell-to-cell contact via plasmids or sex pili. Plasmids are small extrachromosomal DNA molecules that can replicate independently and be transferred between bacteria. Episomes can exist independently or integrate into chromosomes, and include plasmids, viruses, and transposons.
Gene technology
The document outlines the steps involved in genetically engineering bacteria to produce human insulin. These include identifying and obtaining the insulin gene from human pancreas cells, inserting the gene into a bacterial plasmid vector, transforming the vector into host bacteria, selecting transformed bacteria using antibiotic resistance markers, and cloning the bacteria to mass produce human insulin. The document also discusses some advantages of producing insulin through genetic engineering compared to extracting it from animals.
Pac vector ppt
This document discusses PAC (P1 derived artificial chromosome) vectors. PAC vectors are derived from bacteriophage P1 and can accommodate DNA fragments up to 85-100 kb in size, larger than cosmids but smaller than YAC vectors. PAC vectors contain a packaging site (pac) and loxP sites that allow for in vitro packaging of recombinant DNA into phage particles and subsequent circularization of the DNA upon infection of a host E. coli cell expressing Cre recombinase. PAC vectors are useful for constructing genomic libraries from various organisms.
Molecular mapping
Genetic mapping is based on recombination frequencies between genetic loci during meiosis. Physical mapping determines the actual distances in base pairs between sequences on a chromosome using overlapping DNA fragments. Before whole genome sequencing, physical maps were created using techniques like restriction mapping of large-insert clones, probing genomic libraries with end fragments, and chromosome walking to build contigs of overlapping sequences. This allowed sequencing of individual fragments which could then be assembled into a complete genome sequence.
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rDNA Technology-Biosafety Regulations and Guidelines
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Similar to Genetic engineering
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.
General-Biology-2___Recombinant-DNA.pptx
Lourdes conducted research that led to the characterization of over 50 peptides from fish-hunting snail venom and the use of conotoxins to study the human brain. Her discoveries impacted neuroscience as conotoxins continue to be used to examine brain activity.
Ramirez focused on studying the genetics of native plants like coconuts and rice in the Philippines. Her research empowered agricultural scientists and farmers by advancing plant breeding and genetics. She pioneered genetics instruction and was called the "Mother of Cell."
Recombinant DNA technology has revolutionized cell study. Advances in manipulating DNA allowed combining techniques such that researchers can now isolate specific genes to precisely study their structures and functions.
Recombinant dna technology
Recombinant DNA technology involves transferring genes between organisms using artificial means. It works by combining DNA from different sources into a single molecule. The process involves generating DNA fragments, inserting the fragments into vectors, introducing the vectors into host cells, and selecting clones containing the recombinant DNA. Common tools used include restriction enzymes to cut DNA, vectors like plasmids to carry DNA, bacterial hosts like E. coli, and techniques like transformation and selection to introduce and identify recombinant DNA. Applications include analyzing gene structure, producing pharmaceuticals, genetically modified organisms, and gene therapy.
Biotechnology Ap
1. The document discusses genetic engineering and biotechnology, including techniques like recombinant DNA, gene cloning, PCR, and cloning organisms.
2. Key terms defined include genetic engineering, biotechnology, recombinant DNA, gene cloning, restriction enzymes, and plasmids.
3. Recombinant DNA is made by cutting DNA with restriction enzymes and ligating pieces into plasmids, which are then inserted into bacteria.
RECOMBINANT DNA TECHNOLOGY
The document discusses genetic engineering techniques. It describes the stages of gene cloning which include generating DNA fragments, inserting them into a vector, introducing the vector into host cells, and selecting clones. It also discusses various molecular tools used in genetic engineering like restriction endonucleases, vectors, host cells, and methods of gene transfer.
Biotechnology Principles and Processes
The document discusses biotechnology and recombinant DNA technology. It defines biotechnology as using organisms or enzymes from organisms to produce useful products. Recombinant DNA technology involves isolating DNA, fragmenting it with enzymes, inserting fragments into vectors, transforming host cells, and culturing the cells to multiply the DNA. The basic steps are isolating a gene, inserting it into a vector, introducing the vector into a host cell, and using the host to generate multiple copies of the gene.
Gene cloning
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 Resistance
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
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
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.
Role of r dna technology
Recombinant DNA technology uses restriction enzymes and DNA ligase to cut and join DNA fragments from different sources to construct recombinant DNA molecules. This technique was discovered in the 1970s and has since been used to develop transgenic plants with improved traits like higher yield, increased stress and pest resistance, and the ability to produce valuable pharmaceuticals. Some key applications include producing human insulin and anemia treatments, developing herbicide and insect resistant crop varieties, and engineering disease resistance in plants. Recombinant DNA technology is now widely used in agriculture and has contributed to over 70% of foods in supermarkets coming from genetically modified crops.
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.
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.
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.
Cloning dna f inal
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.
Plasmids as vectors
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 and biotechnology 2016
This document discusses genetic engineering and biotechnology. It begins by defining genetic engineering as the manipulation of genes, usually outside an organism's natural reproductive process. It then discusses chromosomes and mutations, including examples of chromosome numbers in different species. Techniques for genetic engineering are explained, such as restriction enzymes and bacterial transformation. Applications include creating transgenic bacteria, plants, and animals. Ethical issues related to genetic engineering are also reviewed.
Basics of molecular cloning
This document provides an overview of cloning techniques including restriction enzymes, joining DNA fragments, plasmid vectors, transforming bacteria, and expressing cloned genes. Key points include:
- Restriction enzymes cut DNA at specific sites and were first isolated in 1970, allowing scientists to cut and join DNA fragments.
- In 1972, Berg et al. created the first recombinant DNA molecule by joining fragments of simian virus 40 and lambda phage DNA.
- Boyer and Cohen later isolated the lacZ gene using restriction enzymes and plasmids, demonstrating genes could be amplified in bacteria.
- Plasmids are commonly used as cloning vectors, with features like antibiotic resistance and multiple cloning sites to insert DNA fragments.
Biotechnology: Principles and Processes Class XII Chapter 11.pptx
Highly descriptive and illustrative presentation based on Biotechnology chapter 11 of NCERT class XII.
This is an important topic especially from biological research point of view.
This is to help students thoroughly understand the topic for exams as well as for future practical applications.
Biotechnology
The document discusses the principles and processes of biotechnology. It describes two core techniques that enabled modern biotechnology: genetic engineering and maintaining sterile environments for microbial growth. It then discusses various tools used in recombinant DNA technology, including restriction enzymes, vectors, competent hosts, PCR amplification, and downstream processing to obtain recombinant products.
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Biotechnology: Principles and Processes Class XII Chapter 11.pptx
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A comparison between organic and conventionally grown carrots prepared on cut...
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Metabolism a
Microbial metabolism involves chemical reactions and workings that allow cells to obtain energy and materials for growth. These reactions are catalyzed by enzymes, which increase reaction rates and lower energy requirements. Adenosine triphosphate (ATP) is the central energy currency that powers cellular reactions but must be continually regenerated through metabolism. Enzymes exhibit high substrate specificity and determine which reactions occur in a cell's metabolism. Metabolic pathways involve interconnected enzyme-catalyzed steps that allow cells to break down nutrients and synthesize cellular components.
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Introduction structure
This document provides an overview of microbiology concepts including:
1) It lists common human pathogens and notes that 61% are zoonotic.
2) It discusses the office hours, labs, and lectures for a microbiology course.
3) It summarizes key discoveries and advances in microbiology, infectious disease, and molecular biology from the 18th century to present.
Growth b
Microbial growth is defined as the increase in the number of cells through cell division, rather than increases in individual cell size. Microbes undergo several growth phases as they multiply, including lag, log, stationary, and death phases, which can be modeled using growth curves. There are various methods to quantify microbial growth, including counting colony forming units, most probable number calculations, and direct microscopic counts. Microbial growth is influenced by various environmental factors such as temperature, pH, oxygen levels, and nutrient availability.
Growth a
The document discusses microbial growth and factors that influence bacterial growth rates. It describes the bacterial growth curve, which consists of lag, exponential, stationary and death phases. The exponential phase is characterized by continuous cell division at maximum rates. Temperature, pH, oxygen levels and water activity are some of the key physical and chemical factors that affect microbial growth rates.
Genetics c
Bacterial cells regulate gene expression in response to environmental changes through transcriptional control mechanisms. The lac and arg operons are two examples that are regulated in Escherichia coli. The lac operon encodes enzymes for lactose metabolism and is induced in the presence of lactose. The arg operon encodes enzymes for arginine biosynthesis and is repressed in the presence of arginine. Operons allow functionally related genes to be coordinately regulated at the level of transcription through repressors and inducers binding to operator sites on the DNA.
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The document discusses several topics related to DNA and genetics including:
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- The genetic code corresponds to DNA base sequences specifying amino acid sequences in polypeptides. Codons and the wobble property are discussed.
- Mutations can be spontaneous or induced by mutagens and cause changes in biochemistry and phenotypes of cells. Different types of mutations like point mutations and frameshifts are described.
- Methods for selecting, detecting, and isolating mutants in microbial genetics experiments are outlined.
- Bacterial plasmids, conjugation, curing of plasmids, and antibiotic resistance factors are
Genetics appt
Bacterial genetics involves three main mechanisms of horizontal gene transfer - transformation, transduction, and conjugation. These mechanisms allow bacteria to acquire new genetic material from other bacteria to help them survive in changing environments. Mutation also contributes to genetic variation in bacteria and usually involves changes to single genes, while gene transfer can involve simultaneous transfer of multiple genes.
Disinfectants b
The document discusses various methods for controlling microorganisms, including physical methods like heat, filtration, and radiation, as well as chemical agents. It provides definitions for key terms like sterilization, disinfection, and sanitization. For physical heat methods, it describes moist heat sterilization techniques like boiling, autoclaving, pasteurization, and ultrahigh-temperature processing. It also covers factors that influence the effectiveness of antimicrobial agents and calculations using D-values and z-values to determine sterilization times.
Disinfectants a ppt
1. The document discusses various methods for disinfecting and controlling microbial growth, including physical methods like heat and radiation, and chemical methods like disinfectants, antiseptics, and other antimicrobial agents.
2. It describes the mechanisms of action for different antimicrobial agents like phenolics, iodophores, alcohols, aldehydes, and oxidizing agents which act by damaging cell membranes, proteins, and nucleic acids.
3. The effectiveness of disinfection depends on factors like concentration and contact time of the antimicrobial agent, amount of organic matter present, and characteristics of the microbes being targeted.
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Antibiotics ppt
- Antibiotics selectively target microbial processes without harming human host cells. Proper antibiotic use and hand hygiene have improved patient outcomes.
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- Antibiotics work by inhibiting bacterial cell wall, protein, or nucleic acid synthesis. However, antibiotic resistance has emerged through various mechanisms and poses a growing challenge.
Nutrition
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- 5. Restriction enzymes Q & A In 1970, Hamilton Smith isolated HindIII (1st restriction enzyme well characterized and used for DNA cloning), which comes from Haemophilus influenzae . They are named based on genus & species of bacteria it was isolated from. (EcoRI = Escherichia coli , RY13). They cut DNA by cleaving phosphodiester bonds (in sugar-phosphate backbone) that join adjacent nucleotides Which was the first one well understood? How are they named? How do they work?
- 12. Vector Features Modern plasmid DNA cloning vectors usually consider 6 desirable features: 1. Size (must be small enough to separate easily) 2. Origin of replication (ori) - DNA sequence at which replication is initiated 3. Multiple cloning site (MCS) - a stretch of DNA with recognition sequences for common restriction enzymes (Engineered into plasmid so that digestion does not result in loss of DNA fragment)
- 18. Vector Features 5. RNA polymerase promoter sequences - place where RNA polymerase binds to begin transcription 6. DNA sequencing primer sequences - known sequence that allows sequencing of cloned DNA fragments that have been inserted into the plasmid