江浙蝮蛇蛇毒蛋白C激活因子基因的克隆及测序

Cloning and sequencing of protein C activator gene from Agkistrodon blomhoffii brevieaudus.

从江浙蝮蛇毒腺中抽提总RNA,RT—PCR进行体外扩增,获得江浙蝮蛇蛇毒蛋白C激活因子基因,克隆至pGEX-5X-3载体中.对3个重组克隆分别作DNA全序列分析.通过遗传密码推导出相应的氨基酸序列.与其它已知的丝氧酸蛋白酶型蛇毒蛋白的氨基酸作比较,其中许多位置上氨基酸有很强的同源性.该基因的成功克隆,不仅推导出江浙蝮蛇蛇毒蛋白C激活因子的蛋白质序列,也为进一步开展江浙蝮蛇蛇毒蛋白C激活因子蛋白质工程的研究工作打下良好的基础.

Total RNA was extracted from the gland of the snake Agkistrodon blomhoffii hrevicaudus. The protein C activator gene then was amplified by RT-PCR and cloned into the pGEX-5x-3 vector. Its nucleotide sequence was determined by analyzing the full DNA sequence of three colonies containing the protein C activator gene. The deduced gene amino acid sequence is analogous to other serine proteases from snakes. The successful cloning of the protein C activator gene not only made the determination of its total amino acid sequence possible, but also provided a good basis for further research work in the protein engineering of protein C activator from snakes.