ANIMAL CLONING VECTOR

Sep 8, 20159 likes5,783 views

This document is a collection of photo credits from various photographers and artists. It includes 10 different photo credits from photographers and artists with names like EverJean, Ajith, eviltomthai, ImAges ImprObables, UweBKK, @Doug88888, saigneurdeguerre, Ice Blade, rgmcfadden, and Birth Into Being. The document ends by encouraging the reader to create their own presentation.

cDNA is produced from mRNA found in the nucleus and contains only the expressed genes of an organism. To create a cDNA library, mRNA is first extracted and purified from a cell, then reverse transcribed into cDNA using an oligo-dT primer that binds to the poly-A tail. The resulting single-stranded cDNA is converted into double-stranded DNA and cloned into plasmids in bacteria. cDNA libraries are useful for reproducing eukaryotic genomes without introns, expressing eukaryotic genes in prokaryotes, discovering novel genes, and studying alternative splicing in different cells.

Shuttle vector - a plasmid vector used in rDNA technology. neeru02

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Shuttle vectors are constructed so that they can propagate in both eukaryotic and prokaryotic cells. They contain two origins of replication - one for each host cell type. YEp13 is an example of a shuttle vector that can replicate in yeast (eukaryotic) and E. coli (prokaryotic) cells. It contains sequences from the 2 micron yeast plasmid, including the LEU2 gene as a selectable marker, as well as sequences from the pBR322 plasmid which allow replication in E. coli. Shuttle vectors allow initial cloning experiments and selection to be done in E. coli before introducing the recombinant DNA into yeast cells.

ISOLATION AND DETECTION OF MUTANTSSudhir Gupta

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This document describes several methods for isolating and detecting bacterial mutants, including replica plating, penicillin enrichment, and the Ames test. Replica plating involves transferring bacterial colonies from a "master plate" to replica plates with and without an amino acid to identify auxotrophic mutants. The penicillin enrichment technique also detects auxotrophs. The Ames test uses Salmonella strains to evaluate whether a chemical is mutagenic based on its ability to increase histidine revertants, indicating DNA damage. Chromogenic substrates can identify mutants unable to utilize substances like lactose due to lacking the enzyme beta-galactosidase.

Phi x 174 phage.qamrunnisashaikh1997

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Retroviral VectorsMaryam Shakeel

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Retroviral vectors are derived from wild type retroviruses like Moloney murine leukemia virus. They are engineered to carry foreign genes into target cells. The vectors contain cis-acting elements from the viral genome like LTRs and packaging signals but lack the trans-acting genes gag, pol, and env. This prevents replication but allows the vector and its gene of interest to integrate stably into the host cell genome. Retroviral vectors show promise for gene therapy applications but also have limitations like requiring actively dividing cells for transduction and potential risks of insertional mutagenesis leading to cancer.

Media formulationeswar1810

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This document discusses the key components required for microbial growth and fermentation, including carbon, nitrogen, minerals, vitamins and oxygen. It outlines the goals of optimizing fermentation media to maximize product yield while minimizing undesirable byproducts. Finally, it examines various carbon sources, nitrogen sources, minerals, trace elements and antifoaming agents used in fermentation media formulation.

Bacteriophage T4 and Bacteriophage lambda HARINATHA REDDY ASWARTHAGARI

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This document discusses two types of bacteriophages - T4 and Lambda phages. It describes their structure, genetic material, life cycles and infection processes. T4 phage infects E. coli and has a long helical tail and base plate with spikes that attach to host cells. Lambda phage also infects E. coli and can undergo lysogenic or lytic cycles. It has genes that determine which cycle occurs based on host cell conditions. The document provides detailed information on the structure, replication, and infection mechanisms of these two important bacteriophages.

Phage vector bacteriophagedennisVARGHESE10

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Strain improvement techniquerekha sharma

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Genomic libraryAman Ullah

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The document discusses genomic libraries. A genomic library contains DNA fragments representing the entire genome of an organism. It is constructed by inserting fragments of partially digested genomic DNA into vectors like plasmids or bacteriophages, which are then introduced into bacteria or yeast. The bacteria replicate and produce clones containing fragments of the original genome, theoretically representing all its sequences. Genomic libraries allow researchers to explore genomic structure and function, map genomes to identify gene locations, and clone DNA segments.

Gene transfer in bacteriakartik1403

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This document provides an overview of gene transfer in bacteria through three main methods: conjugation, transformation, and transduction. Conjugation involves the transfer of genetic material between bacteria via cell-to-cell contact through sex pili. Transformation refers to the uptake of naked DNA by competent bacterial cells. Transduction is the transfer of DNA from one bacterium to another via bacteriophage. Each method is described in 1-2 paragraphs detailing its history of discovery and basic mechanisms.

Enzyme engineering by tamizhTamizh Muhilan

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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.

Blue white screening LisaSam11

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This document describes the blue-white screening technique for identifying recombinant bacteria. Blue-white screening relies on the activity of β-galactosidase, an enzyme in E. coli that cleaves lactose. Plasmid vectors used in cloning carry a fragment of the lacZ gene. When the plasmid inserts foreign DNA at the multiple cloning site, it disrupts lacZ and prevents complementation, so the bacteria cannot metabolize lactose and appear white on an indicator plate. If no DNA inserts or inserts elsewhere, complementation occurs and bacteria appear blue. The technique allows rapid identification of bacteria that took up recombinant plasmid.

Biosynthesis of peptidoglycanKARTHIK REDDY C A

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Peptidoglycan is a polymer that forms the cell wall of bacteria and provides structural strength. It consists of alternating sugars (N-acetylglucosamine and N-acetylmuramic acid) linked together into a mesh-like layer with cross-linking peptide chains. Peptidoglycan is synthesized through a two-stage process where sugar nucleotides and amino acids are assembled into a lipid carrier on the inner membrane before being polymerized into the cell wall. Several antibiotics are able to inhibit peptidoglycan synthesis by binding to enzymes involved in cross-linking, preventing cell wall assembly and causing bacterial cell death.

M13 phagePrakashL12

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TransfectionAchyut Bora

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The document discusses various methods of transfection in animals. Transfection is the process of introducing nucleic acids into eukaryotic cells. It describes viral transfection using bacteria like Agrobacterium tumefaciens and viruses. Non-viral methods include chemical transfection using calcium phosphate, liposomes, polyamines. Mechanical transfection employs microinjection or particle bombardment. Common chemical methods are calcium phosphate precipitation, polyplexes, and liposomes/lipoplexes. Viruses used are retroviruses, adenoviruses, adeno-associated viruses. Bacterial and viral vectors allow for integration into the host genome while chemical and mechanical are often transient.

Reporter gene[2]Dilip Pandya

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Reporter genes are genes that produce easily detectable and quantifiable proteins to track the expression of other genes. Common reporter genes include GFP, luciferase, CAT, and β-galactosidase. Reporter genes are used to study gene expression patterns, monitor plant transformation, and study regulatory elements. There are two main types of reporter genes - scorable markers, which produce a quantifiable phenotype, and selectable markers, which allow cells to survive under selective conditions using antibiotic resistance.

Abzymesjeeva raj

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Abzymes, also known as catalytic antibodies, are monoclonal antibodies that exhibit enzymatic activity. They are able to bind to transition states of enzyme-catalyzed reactions with high specificity and affinity, stabilizing the transition state and increasing reaction rates. Abzymes can be artificially produced by immunizing animals with transition state analogs of reactions. They have potential applications in drug development, cancer treatment, and developing therapies for viral infections like HIV. Researchers have engineered an abzyme that can degrade an essential region of the HIV envelope protein, rendering the virus unable to infect cells.

Sv 40Yashasvi Kumar Singh

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Simian virus 40 (SV40) is a DNA virus that can cause tumors in monkeys and humans, and it was first identified as a contaminant in polio vaccines in the 1960s. SV40 has been widely used as a cloning vector due to its ability to efficiently deliver genes into a variety of cells without killing the host cell or eliciting an immune response. Future research prospects for SV40 vectors include developing recombinant versions for gene transfer applications and furthering understanding of related retroviruses.

Taq dna polymerase - enzyme used in PCR amplification technologyneeru02

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Taq polymerase is a thermostable DNA polymerase enzyme isolated from the bacterium Thermus aquaticus. It is used for polymerase chain reaction (PCR) because it can withstand the high temperatures needed to denature DNA during PCR cycling. Taq polymerase has optimal activity between 75-80°C and can replicate 1000 base pair DNA strands in under 10 seconds at 72°C. However, it lacks proofreading ability, resulting in a relatively high error rate of around 1 in 9,000 nucleotides.

Shuttle vectorkishoreGupta17

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1. Yeast plasmids like the 2 micron circle have been extensively studied and developed into yeast cloning vectors. 2. Shuttle vectors like YEp vectors contain selectable marker genes like LEU2 and bacterial plasmid origins of replication like pBR322, allowing them to replicate in both E. coli and yeast. 3. The 2 micron circle is a 6kb endogenous yeast plasmid that replicates autonomously through an ARS sequence and is maintained at 50-100 copies per cell.

Isolation of mutantsshveta arya

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Animal viral based vectorsPinky Premmy

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Viral vectors are efficient tools for gene delivery due to viruses' ability to transfer DNA into host cells. The document discusses several types of viral vectors, including adenoviral, adeno-associated, retroviral, lentiviral, and baculovirus vectors. It provides details on the structure and genome organization of different viruses used to create these vectors. The document also explains the process of generating recombinant viral vectors by removing unnecessary viral genes and inserting genes of interest. Viral vectors allow for transient or stable gene expression and are useful for both research and clinical applications such as gene therapy and vaccine development.

LAMBDA PHAGE AND THEIR REPRODUCTION BanumathiKK

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Lambda phage is a bacteriophage that infects E. coli and can undergo either lytic or lysogenic replication cycles. In the lytic cycle, the phage attaches to the bacterial cell and injects its DNA, which is then replicated and structural proteins are synthesized. New virions are assembled and released upon lysis of the host cell. In the lysogenic cycle, the phage DNA circularizes and integrates into the bacterial chromosome to form a prophage. The lysogenic bacteria replicate normally with the prophage until it enters the lytic cycle again. The genetic switch between lytic and lysogenic cycles is regulated by the cro and CI proteins of the phage.

Phagemid and bac vectorsPromila Sheoran

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The document summarizes phagemid and bacterial artificial chromosome (BAC) vectors. It describes that phagemid vectors are plasmids that contain both plasmid and phage origins of replication. Specifically, it discusses the features of pBluescript II phagemid vectors, including their polylinker and RNA polymerase promoter sequences. It also describes how pBluescript II phagemid vectors can produce blue or white colonies depending on insert presence. The document then explains that BAC vectors are low-copy plasmids that can hold up to 300kb DNA fragments. Examples of BAC vectors like pBAC108L and pBeloBAC11 are provided, with details about their replication origin and partitioning functions.

Mutagenesisberciyalgolda1

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transformationIram Khan

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This document discusses bacterial transformation, which is the process by which bacteria take up naked DNA from their environment. There are three main ways bacteria exchange DNA: conjugation, transduction, and transformation. Transformation occurs when bacteria take up free DNA from their surroundings. Some bacteria have the natural ability to undergo transformation, while others can be artificially induced to become competent through chemical treatments or electroporation. The document outlines the key steps and proteins involved in natural and artificial bacterial transformation.

MicroinjectionNikita Narwal

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Viruses as vector, binary, shuttle vectorPromila Sheoran

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This document discusses different types of vectors that can be used for genetic engineering, including animal viruses, plant viruses, retroviruses, shuttle vectors, and binary vectors. It provides details on the structure and use of tobacco mosaic virus (TMV) as a plant viral vector, describing how foreign genes can be stably replicated and spread systemically in plants using this system. It also summarizes key features of yeast episomal plasmids (YEps), retroviral vectors based on murine leukemia virus, and the binary vector system used for plant transformation via Agrobacterium tumefaciens.

VIRAL VECTORS FOR GENE TRANSFERANKUR SHARMA

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