Lecture 19 Chapter 20

Lecture 19 Chapter 20 - PowerPoint Slides for Introduction...

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  1 PowerPoint Slides for Introduction to Genetic Analysis Ninth Edition Anthony J. F. Griffiths, Susan R. Wessler, Richard C.  Lewontin, and Sean B. Carroll CHAPTER 20 Gene Isolation and Manipulation Copyright 2008 © W H Freeman and Company
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  2 Genetic Engineering The ability to isolate (clone) genes and manipulate genes led  to the birth of the new field of genetic engineering.
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  3 Genetic Engineering 1. Important factors 1. Ability to fragment the genome. 1. Restriction enzymes 1. Ability to fuse two different DNA molecules. 1. Complementary base pairing  2. DNA ligase 1. Ability to amplify a DNA region or gene. 1. DNA polymerases 2. Reverse transcriptase Ability to zoom in on a specific DNA region or gene. Molecular Probes
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  4 Gene Manipulation Cloning DNA or copy of RNA Genomic clone  DNA region containing a specific Gene cDNA clone  RNA copy of a specific Gene Genomic DNA library Entire genome of an organims cDNA library DNA copies (cDNA) of all the mRNAs which are expressed in a specific cell type. Chromosome walking.  To clone large chromosomal regions Using one gene as anchor to get to another gene Cloning a disease gene. From disease to Gene Using gene clone for DNA diagnostics. From Gene to disease Probing gene function using genetic engineering.
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  5 Chromosome Walking Use one DNA clone as an anchor and isolate overlapping clones to get  to another place on the same chromosome  Cloning   Clone DNA fragment into  a vector (a DNA molecule  which can replicate  autonomously in a  bacterial host) PCR Amplify DNA fragment directly  using thermostable polymerases  and PCR DNA Cloning In order to study a gene or a specific region in our genome we need to isolate that  region and make a large amounts of it
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  6 DNA Cloning
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  7 Recombinant DNA Technology Major discoveries which led to the birth of genetic engineering  and recombinant DNA technology 1. Restriction enzymes   1. Binds to specific sequences within the DNA molecule. 1. Fragments DNA into smaller pieces. 1. Creates single stranded overhangs (sticky ends). 1. DNA Ligases 1. Can fuse sticky ends created by restriction digested DNA fragments through  complementary base pairing. 1. Able to fuse two different DNA fragments of different origins (human and  bacterial etc.).
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This note was uploaded on 03/15/2009 for the course BIOL 3301 taught by Professor Gunaratne during the Fall '05 term at University of Houston.

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Lecture 19 Chapter 20 - PowerPoint Slides for Introduction...

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