甘蓝型油菜突变体Cr3529抑制性消减文库的构建  

The Construction of the Suppression Subtractive Library of the Brassica napus Mutant Cr3529

作　　者：刘阳[1] 王茂林[1] 邱峰[1] 周云涛[1] 赵云[1] 张帆[2] 

机构地区：[1]四川大学生命科学学院,成都610064 [2]四川省种子管理站,成都610041

出　　处：《四川大学学报（自然科学版）》2005年第5期1029-1032,共4页Journal of Sichuan University(Natural Science Edition)

基　　金：国家自然基金科学资助项目(30170500)

摘　　要：应用抑制性消减杂交(suppression subtractive hybridization,SSH)技术构建了甘蓝型幼叶黄化油菜突变体Cr3529和其野生型3529品系幼叶期的正、反向消减文库.经菌落PCR检测,正向消减文库的有效重组率为55.5%;反向消减文库的有效重组率为76.5%.为进一步克隆消减文库中的差异表达基因和研究这些基因的功能奠定了基础.The forward and backward subtraction libraries of young leaves in chlorophyll- reduced mutant Cr3529 and its wild type 3529 were constructed by suppression subtractive hybridization （SSH）. The express levels of these genes were changed greatly after the suppression subtractive hybridization, that is, they were obviously enriched by SSH. The secondary PCR products of the forward and backward subtraction were cloned, inserted into the pMD18-T vector and transferred into E. coli. These clones in the forward and backward subtraction libraries were verified by colony PCR. The reeombined efficiencies of the forward and backward subtraction libraries were 55.5% and 76.5%, respectively.

关 键 词：抑制消减杂交 消减文库 菌落PCR 

分 类 号：Q81[生物学—生物工程]