鸡BPI氨基端的基因克隆和鉴定

Cloning and identification of chicken′s BPI protein N-terminal fragment

应用RT-PCR技术,参照GenBank报道的预测序列,从鸡多形核白细胞(PMN)mRNA中扩增出杀菌/通透性增加蛋白(BPI)氨基端95个氨基酸的基因片段,并与pMD18-T连接,构建BPI氨基端的基因重组体,进行酶切鉴定和序列测定。结果表明,获得了BPI N端长度为284 bp的基因片段,序列分析证实该片段中有3个点突变,但均不在活性中心。

According to the prediction sequences of GenBank, the gene which encode N-terminal fragment of BPI protein were amplified by RT-PCR from mRNA extracted from chicken polymorphonuclear neutrophils （PMN）. The PCR product was cloned into the pMD18-T vector and the sequence was confirmed by restriction enzyme digestion and dideoxy-mediated chain in termination. The results showed that the sequences are the gene of chicken＇s BPI.