灰飞虱酵母双杂交cDNA文库的构建及分析

摘要

A yeast two-hybrid cDNA library was constructed using the CloneMinerTM cDNA Library Construction Kit. The results showed that the unamplified library consisted of 1.85 × 107 independent clones; the titer of amplified cDNA library was 7.7× l08 cfu/mL, and the recombinant rate was about 97％. The sizes of most inserts were 1 000-1 500 bp in length. Random sequence analysis of ten clones showed that seven of the clones had homologous sequences in GenBank. Clones L2 and L9 were the same as the published sequences. Construction of this yeast two-hybrid cDNA library of Laodelphax striatellus laid the foundation for cloning of functional genes in L.striatellus and studies on L. striatellus-virus interactions.%以灰飞虱(Laodel phax striatellus Fallen)mRNA为起始模板,利用Gateway技术构建了灰飞虱酵母双杂交cDNA文库.经过检测表明:构建的初级cDNA文库的库容量为1.85×10<'7>cfu;扩增文库滴度为7.7×10<'8>cfu/mL,重组率约为97%;扩增文库插入片段主要集中在1 000～1 500 bp之间.随机挑取10个克隆,经测序与GenBank数据库比对结果显示7个克隆具有同源序列,其中L2、L9为已公布的灰飞虱序列.灰飞虱酵母双杂交cDNA文库的构建为克隆全长目的基因及研究灰飞虱与其传播的水稻病毒间的互作奠定了基础.