经大肠杆菌感染的中国家蚕cDNA文库的构建

Establishment cDNA Library of the Silkworm ( Bombyx mori ) from China Infected by E.coli

用建立非表达文库的方法，从大肠杆菌感染后９ｈ的中国家蚕的ｍＲＮＡ合成ｃＤＮＡ片段，重组入噬菌体λｇｔ１０的ＥｃｏＲＩ的位点之间，所构成的基因非表达文库库容量为２×１０６重组子，经大肠杆菌Ｃ６００与Ｃ６００ｈｌｆ菌株平皿测定，重组率为７６％．

cDNA fragments were obtained by in vitro transcription of the mRNA isolated from the fat bodys in the silkworm of China,9 hours after infected by E.coli K12D31. After Eco R I adaptor ligation, the cDNA was directionally ligased with the Eco R I arms of the Lambda λgt10 vector, followed by Lamda DNA Packaging and infected host bacterial strain C600 hlf . According to the phage plaques bright selection,the percentage of recombinant phages are 76%,while the cloning effeciency are high too, and the titer is 2×10 6 pfu/μg.