牦牛肾脏cDNA文库的构建

cDNA Library construction of the yak kidney

目的 :构建牦牛肾脏的cDNA文库 ,为进一步筛选、克隆和表达有益基因做准备。方法 :从牦牛肾脏中提取总mRNA ,经逆转录合成cDNA后 ,以pSPORT1质粒为载体构建cDNA文库。结果 :牦牛肾脏cDNA文库库容量为1×106转化子/μgcDNA ,经E.coliDH5α菌株平板测定 ,重组率为93 %。结论 :经平板鉴定和PCR鉴定表明 ,所构建cDNA文库达到建库要求 。

Objective: To construct the cDNA library of yak kidney.Methods: Non-expression cDNA library of yak kidney was constructed by using a highly efficient and simplified cDNA cloning method for the development of genetically engineered products. After mRNA extracted from the yak kidney,cDNA was obtained by reverse transcription, and then transformed into E.coli DH5α by plasmid pSPORT1.Results: The efficiency of cloning was 1×106pfu/μg cDNA and the percentage of recombinant phages are 93%.Conclusion:The result shows that the library is available for screening target gene and expression of proteins.