蝮蛇毒碱性磷脂酶A_2基因的克隆

从蝮蛇毒腺中抽提总ＲＮＡ．利用人工合成寡核苷酸引物作逆转录，以ｃＤＮＡ为模板进行体外扩增，获得磷脂酶Ａ２（简称ＰＬＡ２）基因，克隆至ｐＢＳ－ｋｓ载体中。通过对３个碱性ＰＬＡ２（简称ＢＰＬＡ２）基因单独克隆分别作ＤＮＡ全序列分析，推导ｐｒｏ－ＢＰＬＡ２由１３８个氨基酸残基构成，与已测定的部分氨基酸序列比较，基本相符。该基因成功的克隆，不仅推导出ＢＰＬＡ２的蛋白质全序列，也为进一步开展蛇毒功能肽蛋白质工程的研究工作打下了良好的基础。

Cloning of the BPLA_2 Gene from Agkistrodon halys Pallas

First Strand cDNA synthesis was primed with synthetic oligonueleotide from total RNA extracted from the gland of the snako Agkistrodon halys Pallas.The pro-BPLA2 gene was then amplified by PCR and cloded into the pBS-KS vector. Its nucleotide sequence has been determined by analyzing the DNA sequence of three colonies containing the pro-BPLA2 gene. The deduced amino acid sequence consists of 138 amino acids and agrees with the Partly known amino acid sequence except for seven amino acid residues. The successful cloning of the BPLA2 gene not only has 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 functional peptides from snakes.