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The study of transposition mechanism and the biology of transposons is an interesting subject in gen

The study of transposition mechanism and the biology of transposons is an interesting subject in gen

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The study of transposition mechanism and the biology of transposons is an interesting subject in genetics but for our current purposes we are going to concentrate on how transposons can be used for bacterial genetic analysis. For this purpose we will focus on the transposon TTTTT which can function in E. coli as well as a wide variety of other bacterial species. The selectable marker in TTTTT is a gene that confers resistance to the antibiotic kanamycin. Thus bacteria without TTTTT are sensitive to kanamycin (Kan s ), whereas bacteria that have TTTTT inserted into the chromosome are resistant to kanamycin (Kan r ).
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Unformatted text preview: To introduce random insertions of TTTTT into the E. coli chromosome we will start with T carried on a special λ phage vector: λ λλ λ P P P – –– :: : . PPPPP allows conditional phage growth. When λ λλ λ P P P phage infect E. coli with an amber suppressor (Su + ) the phage multiply normally, but when λ λλ λ P P P phage infect a nonsuppressing host (Su – ) the phage cannot replicate. i – is a mutation in the λ integrase gene. Phage with this mutation can not integrate into the host chromosome to make a stable prophage. : designates that the λ phage carries an inserted copy of TTTTT ....
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