尖吻蝮蛇去整合素基因克隆与功能分析  

Cloning and biological characteristic of a novel disintegrin gene from Deinagkistrodon acutus

作　　者：姜升阳[1] 张艳宇[1] 马平[1] 吕丽萍[1] 周锡鹏[1] 许金波[1] 

机构地区：[1]军事医学科学院野战输血研究所,北京100850

出　　处：《军事医学科学院院刊》2006年第1期51-54,共4页Bulletin of the Academy of Military Medical Sciences

基　　金：国家自然科学基金资助(30400272)

摘　　要：目的:获得尖吻蝮蛇去整合素基因,并分析重组蛋白生物学活性。方法:提取尖吻蝮蛇毒腺组织总RNA,经RT-PCR获得去整合素基因片段,将该去整合素基因片段重组入pMAL-p2x载体中诱导表达,以直链淀粉树脂柱亲和纯化,并检测融合蛋白的抑制血小板聚集活性。结果:该片段编码的氨基酸序列中含有去整合素保守的RGD三肽序列及12个Cys残基,与GenBank中其他蛇毒去整合素氨基酸序列最高同源性为86%,重组融合蛋白具有抑制ADP诱导的血小板聚集活性。结论:发现了一个新的去整合素基因,该基因编码蛋白具有抑制血小板聚集的功能。Objective; To get disintegrin gene sequence of Deinagkistrodon acutus and analyze its protein activity for developing anticoagulant drugs. Methods：Total RNA was extracted from Deinagkistrodon acutus snake venom gland, and the disintegrin gene was amplified by BT-PCR. The prokaryotic expressing vector was constructed, induced by IPTG, and the recombinant protein was expressed. The activity of recombinant protein for inhibiting platelet aggregation in vitro was primarily analyzed after the protein was purified by amylose resin, Results ：The sequence of amino acids encoded by the gene contained a RGD tri-peptide sequence and 12 eysteines which were eoneervative in position and amount in disintegrim. There was at maximum 86% homology between the new sequence of amino acids and other disintegrins in GenBank. The recombinant protein has the ability to inhibit platelet aggregation induced by ADP. Conclusion：A new disintegrin gene was discovered, and the protein encoded by this gene has the bioaetivity of inhibiting platelet aggregation.

关 键 词：尖吻蝮蛇 去整合素 基因工程 血小板聚集 

分 类 号：R96[医药卫生—药理学]