版纳微型猪近交系外周血淋巴细胞cDNA文库的构建

CONSTRUCTION OF CDNA LIBRARY FROM PERIPHERAL LYMPHOCYTE OF CHINESE BANNA MINIPIG INBRED LINE

[目的]构建版纳微型猪近交系(BMI)外周血淋巴细胞中cDNA文库。[方法]取新鲜BMI外周血,分离淋巴细胞,并进行体外诱导培养,提取总RNA,纯化mRNA。按照SMART cDNA library construction kit提供的方法构建BMI外周血淋巴细胞cDNA初始文库,进行滴度测定及文库扩增。[结果]初始文库和扩增文库的滴度分别为1.1×106pfu/ml和2.5×109pfu/ml,重组率分别为98.2%和89.1%,插入片段平均长度大于1200bp。[结论]成功地构建了BMI外周血淋巴细胞cDNA文库。

[ Objective] To construct a peripheral lymphocyte cDNA library of Chinese Banna Minipig Inbred Line （BMI）. [Methods] We extracted total RNA and purified mRNA of peripheral lymphocyte from one of the 16th inbred generation of BMI JS 151 substrain. Then a peripheral lymphocyte cDNA library was constructed according to the protocol from SMART cDNA library construction kit. [ Results] The titers of the primary and amplified libraries were 1.0×10^6 pfu/ml and 2.5×10^9 pfu/ml respectively. The percentages of recombinants were 98.2% in primary library and 89.1% in amplified library. The average length of inserts was more than 1 200 bp. [Gonclution] We successfully construct a peripheral lymphocyte cDNA library of BMI.