Lecture 40 and 41 DNA technology

Lecture 40 and 41 DNA technology - Lectur e 40 AN D 41: DN...

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Unformatted text preview: Lectur e 40 AN D 41: DN A technology - Basic pr inciples and techniques Campbell, CH APTER 20 Lear ning objectives 1. Be able to explain the basic steps involved in generating and amplifying a recombinant clone. 2. Define the role of each of the following in recombinant DNA technology: ligase, restriction endonuclease, transformation, selection, plasmid. 3. Understand the principles of gel electrophoresis and be able to explain its applications. 4. Explain how nucleic acid hybridization is used to identify particular nucleic acid molecules. 5. Understand the principles of Southern blotting and be able to explain its applications. 6. Understand the PCR reaction and describe some of its applications. 7. Define cDNA and its importance during the expression of eukaryotic genes in bacteria. Do all of the end of chapter problems (pp 424, 425) Advances in DNA technology has allowed biotechnological applications , including introduction of genes into hosts for large scale production of pharmaceutical products (e.g. human hormones, vaccines, other proteins). medical applications that allow rapid diagnosis of genetic and other diseases agricultural applications, including production of disease-resistant plants production of transgenic organisms (mice, sheep, chickens, goats, used for basic research or as production factories) cloning of mammals. environmental cleanup using genetically modified bacteria that can degrade potentially toxic waste genomic sequencing of thousands of organisms Forensic application- analyses of biological crime samples that provide evidence for or against guilt/ parental disputes These methods form part of genetic engineer ing , the direct manipulation of genes for practical purposes. To study a particular gene, well-defined portions of a chromosome containing the gene of interest needs to be isolated, amplified, and char acter ized . DNA Cloning Recombinant DNA (aka DNA cloning) is the cutting and mixing of DNA from two different sources. I n practice, a particular fragment of DNA is isolated from one source, then pasted into a vector (usually a plasmid or virus) that can replicate many times when introduced in an appropriate host cell (usually in bacteria). This allows that fragment of DNA DNA to be amplified at quantities sufficient for further analysis. High copy expression of protein- encoding DNA in a suitable host also allows the protein to be produced at high quantities. TH E GOAL OF DN A CLON I N G I S TO AMPLI FY A PARTI CULAR DN A SEQUEN CE Centr al to all r ecombinant DN A technology ar e the following pr inciples: 1. E nzymes recognize and modify nucleic acid in vitro with high specificity. Restriction endonucleases cleave DNA at specific sequences, ligases glue DNA molecules together, and polymerases synthesize DNA or RNA....
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This note was uploaded on 11/14/2010 for the course CHE 131 taught by Professor Kerber during the Spring '08 term at SUNY Stony Brook.

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Lecture 40 and 41 DNA technology - Lectur e 40 AN D 41: DN...

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