中国林蛙皮cDNA文库的构建

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

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.