鼠疫耶尔森氏菌LcrV基因的克隆及序列分析

Cloning and sequencing of LcrV-gene of Yersinia pestis

为了研究鼠疫耶尔森氏菌 (Y .pestis)保护性抗原V蛋白 ,从基因库中查得Y .pestisLcrV基因DNA序列 ,针对序列设计合成了一对PCR扩增引物 ,以本所保存的Y .pestis菌种为模板进行基因扩增 ,结果获得长约 980bp的DNA片段。将扩增产物回收纯化 ,克隆至pGEM T载体 ,构建重组载体pGEM T/ypV ,经过PCR ,酶切鉴定 ,并对pGEM T/ypV中的V基因片段进行测序 ,分析测序结果与已知序列相同 。

To study the protective antigen V protein of Yersinia pestis, a pair of primers for PCR were designed according to published DNA sequence of Y. pestis LcrV gene and synthesized. A DNA fragment about 980 bp length was gotten through PCR amplification. After recycled and purified, the DNA fragment was ligated with pGEM?T Vector and recombined vector pGEM?T/ypV was constructed. The pGEM?T/ypV plasmid was then identified by PCR, enzyme digestion and sequencing, and the sequence was the same as the published sequence.