中国林蛙皮cDNA文库的构建  

Construction of cDNA Library from Skin of Rana temporaria chensinensis,David

作　　者：周杰[1] 孟庆繁[1] 丁天兵[1] 逯家辉[1] 田晓乐[1] 李青山[1] 滕利荣[1] 

机构地区：[1]吉林大学生命科学学院,吉林长春130012

出　　处：《生物技术通讯》2007年第5期792-793,共2页Letters in Biotechnology

基　　金：吉林省科技厅资助项目(200305-50-6)

摘　　要：目的:为研究林蛙皮抗菌肽,筛选、克隆及表达相关抗菌功能基因,构建林蛙皮cDNA文库。方法:提取林蛙皮总RNA后,利用v-gene纯化试剂盒纯化mRNA,RT-PCR合成双链cDNA;取10ng/μLcDNA按照不同比例连接到λ噬菌体载体,以选择最佳连接比例。结果:获得克隆总数为1.2×105的林蛙皮cDNA文库,重组率为99.4%。结论:所建立的cDNA文库可用于进一步筛选、克隆抗菌功能新基因。Objective： To construct a high quality cDNA library from the skin of Rana temporaria chensinensis, David. Methods： After exaction of total RNA, mRNA was purified by using v-gene mRNA purified kit. The cDNA was synthesized through reverse transcription PCR, after that the double cDNA was ligated to λ phage vector. Results： The directional library was confirmed to be about 1.2×10^5 independent clones in which the percentage of recombinant clones was 99.4%. Conclusion： The constructed eDNA library can be used for further screening and cloning of new antimicrobial gene in Rana temporaria chensinensis, David.

关 键 词：中国林蛙 CDNA文库 文库滴度 重组率 

分 类 号：Q78[生物学—分子生物学]