长白猪甘露聚糖结合凝集素A基因的 克隆与原核表达

从长白猪肝脏细胞中提取总RNA，通过RT-PCR法扩增pMBL-A基因，与pMD18-T载体连接进行TA克隆，得到猪甘露聚糖结合凝集素A（porcine mannan-binding lectin A，pMBL-A）基因。再亚克隆到pPROEXHTb表达载体上，构建重组表达质粒。将阳性重组表达质粒转化到大肠杆菌中诱导表达重组pMBL-A蛋白，通过SDS-PAGE检测蛋白的表达情况。结果成功扩增得到包括完整开放读码框长为875bp的全长cDNA片段，并原核表达了重组蛋白，为进一步研究该蛋白的遗传特征以及为猪遗传育种方面的研究提供依据。

Cloning and Prokaryotic Expression of Porcine Mannan-binding Lectin A Gene

Total RNA was extracted from liver cells of landrace, pMBL-A gene was amplified by RT-PCR. It was cloned to pMD18-T vector, then the mannan-binding lectin A gene of landrace was obtained. It was subcloned to pPROEXHTb vector for expression, recombinant expression plasmid was constructed. The recombinant expression plasmid was transformed into E.coli and was induced to express recombinant protein of pMBL-A. The expressed protein was identified by SDS-PAGE. The cDNA of pMBL-A gene was successfully cloned, its full-length was 875bp including the whole ORF. Recombination protein was pronuleus expressed, it provides affordance for the research of genetic character of the gene and genetic breeding of pigs.