紫茎泽兰cDNA文库的构建及RuBP羧化酶基因的筛选

CONSTRUCTION OF cDNA LIBRARY AND RuBP CARBOXYLASE GENE SCREENING OF Eupatorium adenophorum

采用SMART技术首次构建了高质量的紫茎泽兰cDNA文库。经过涂平板测定和酶切反应鉴定表明,原始文库的滴度为1.23×107cfu/ml,重组率达98%,插入片段平均大小在900bp左右,采用涂平板均匀扩增得到的扩增文库滴度达到了1×1012cfu,覆盖率达99%以上。利用该文库筛选得到了紫茎泽兰核酮糖二磷酸羧化酶(RuBP羧化酶)cDNA片段。

cDNA library of Eupatorium adenophorum was constructed by using SMART system. According to the evaluation, the constructed library had a high titer of 1.23×10~7 cfu/ml, an average insert fragments size was about 900bp and 98% recombinant percentage. The titer of amplified library was 1×10~ 12 cfu by amplification of colonies, the coverage of the library was more than 99%.The cDNA fragment of ribulose diphosphatecarboxylase （RuBP） was obtained by screening this cDNA library.