用基于galK的同源重组的方法构建arc基因表达增强的载体

Construct Vector with Enhanced Arc Expression by GalK-based Recombineering

目的:利用同源重组的方法,将arcBAC(bacteria artificial chromosome)启动子区域的SRE(serumre-sponse element)改造成Gal4结合序列,以此通过Gal4/UAS系统增强arc基因表达。方法:在大肠杆菌中利用依赖于噬菌体基因的同源重组,通过galK正负筛选的方法改造arcBAC。结果:成功将arc启动子区域的SRE改造成Gal4结合序列,并且没有影响arc基因的其他序列。结论:成功改造BAC,可以将此BAC用在Gal4/UAS系统增强arc基因表达的实验中。

Objective：In order to enhance arc expression using Gal4/UAS system,we replace SRE with Gal4 binding site（UAS）.Methods：arc BAC is modified via galK positive/counterselection in the context of recombineering.Results：The SRE site is replaced by Gal4 binding sequence without introducing any other DNA sequences.Conclusion：arc BAC is reconstructed successfully.