Nissley D V, Lindh F G, Fennewald M A
Department of Microbiology and Immunology, University of Health Sciences, Chicago Medical School, IL 60064.
J Mol Biol. 1990 Jun 20;213(4):671-6. doi: 10.1016/S0022-2836(05)80254-2.
The transposase protein and the terminal inverted repeat sequences of the prokaryotic transposon Tn3 are essential for transposition. In order to determine the sequences within the inverted repeat necessary for transposition and interaction with transposase, we have constructed a series of mini-Tn3s in which specific mutations have been introduced into the inverted repeats. The effects of these mutations on transposition have been assayed in vivo using a mating-out transposition assay. Several single base-pair mutations within the transposase binding site reduce transposition frequency. Mutations that affect transposition show a greater effect when present in both inverted repeats than when present in only one inverted repeat.
原核转座子Tn3的转座酶蛋白和末端反向重复序列对于转座至关重要。为了确定反向重复序列中对于转座以及与转座酶相互作用所必需的序列,我们构建了一系列微型Tn3,其中特定突变已被引入到反向重复序列中。这些突变对转座的影响已在体内通过交配输出转座试验进行了测定。转座酶结合位点内的几个单碱基对突变降低了转座频率。影响转座的突变在两个反向重复序列中都存在时比仅在一个反向重复序列中存在时具有更大的影响。