两个标准化cDNA差减文库的构建

CONSTRUCTION OF 2 NORMALIZED cDNA SUBTRACTED LIBRARIES

目的 构建 2个标准化 c DNA差减文库。方法 以淡色库蚊对溴氰菊酯抗药性品系为Tester、敏感性品系为 Driver,进行正向抑制性差减杂交 ;以敏感品系为 Tester、抗性品系为 Driver,进行反向抑制性差减杂交。分别将获取的 2组抑制性差减杂交产物克隆入质粒表达载体 (Pin-Point TMXa- 1T- Vector) ,并以 PCR扩增鉴定插入片段。结果 获得 2个标准化 c DNA差减文库 ,其中抗性相关基因 (抗性品系高表达或特异表达 )文库含 5 2 3个重组子 ,敏感相关基因 (敏感品系高表达或特异表达 )文库含 2 86个重组子。插入片段大小平均约 36 0 bp。结论 所建的

Objective To construct 2 normalized cDNA subtracted libraries. Methods By suppression subtractive hybridization, differentially expressed genes ofCulex pipiens pallensbetween resistant and susceptible strains were got and cloned into PinPoint plasmid vector. ]Results Two normalized cDNA subtracted libraries were obtained including 523 and 286 recombinants respectively. The length of fragments inserted was average 360 bp.Conclusion The 2 constructing subtracted cDNA libraries were suitable for further study.