蓝点马鲛头肾cDNA文库的构建及鉴定

Construction of cDNA Libraries and Identification of Head Kidney from Scomberomorus niphonius

研究运用SMART技术对蓝点马鲛头肾构建了cDNA文库,该文库原始文库滴度为2.26×106pfu/m L,文库容量为1.36×106pfu,符合文库构建的要求。从文库中随机挑选24个单克隆菌,以载体的通用引物进行菌落检测PCR,结果表明:重组率为95.8%,插入片段大小平均为1 000bp。该文库的构建满足了基因的分离与筛选,为进一步研究其相关基因克隆及分子生物学研究奠定基础。

cDNA library of Scomberomorus niphonius head kidney was constracted by SMART technology. The original library titer is 2.26×106pfu/mL,and library capacity is 1.36×106pfu,meeting the requirements of library construction. RandomLy selected 24 monoclonal bacteria from the library,subjected to colony PCR detected by using universal primers. The results showed that recombination rate was 95.8%,an average in-sert size was 1 000bp. Construction of the library meet the isolation and screening of gene,and lay the foundation for further study of melecμlar biology and gene cloning.