Lecture 13 - Lecture 13 Gene Manipulation in Bacteria...

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Lecture 13 Gene Manipulation in Bacteria There is no meiosis in bacteria so special techniques have been worked out for manipulating genes in bacteria so  that mapping experiments, strain construction, and complementation tests can be done.  First, we need a way of getting chromosomal DNA from one cell into another. There are several ways to do this. All  of the methods have in common the use of special extra chromosomal elements for mobilizing chromosomal genes; the  methods differ according to which extra chromosomal element is used.  We will consider a method that uses phage and is known as  Transduction E. coli chromosome is 4.6 x 10 base pairs  Phage  P1 chromosome is 10 base pairs  After infection of E. coli, the phage DNA is replicated by a mechanism known as a “rolling circle” and the phage is  packagedinto phage particles one headfull at a time:   1/300 phage mistakenly packages E. coli chromosome DNA instead of phage DNA.
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Each phage particle will package about 1/50 of the E. coli chromosome. By combining probabilities we see  that about 1/15,000 phage will carry a particular E. coli gene.  A basic transduction experiment to measure the linkage between markers  A and  B is done as follows:  (1) Grow P1 on  A+B+ (2) Infect  A–B– (3) Select for  A+ and then screen for  B+ The idea is that we are looking for the rare cases where some chromosomal DNA carrying gene 
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This note was uploaded on 01/20/2012 for the course GENETICS 380 taught by Professor Glodowski during the Fall '08 term at Rutgers.

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Lecture 13 - Lecture 13 Gene Manipulation in Bacteria...

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