中国长白山乌苏里蝮蛇降纤酶基因克隆和结构分析  

Gene Cloning and Sequence Analysis of Defibrasefrom Gloydius ussuriensis Snake

作　　者：孙德军[1] 闫琪[2] 杨春伟[1] 杨同书[1] 颜伟群[1] 侯立中[1] 

机构地区：[1]吉林大学再生医学科学研究所生物化学教研室,吉林长春130021 [2]长春中医学院

出　　处：《吉林大学学报（医学版）》2003年第2期138-141,共4页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助项目 (39870 375 )

摘　　要：目的 :克隆乌苏里蝮蛇 ( Gloydius ussuriensis)降纤酶 ( Difibrase)基因 ,为用基因工程方法生产降纤酶打下基础。方法 :从乌苏里蝮蛇毒腺制备总 RNA,经 RT- PCR扩增、克隆和顺序测定。结果 :降纤酶 c DNA开框读码序列全长 70 2个核苷酸 ,编码 2 34个氨基酸 ,推测其活性中心为His41、Asp86和 Ser180 ,含有 6对二硫键。结论 :确定乌苏里蝮蛇降纤酶 c DNA顺序 ,推导出其氨基酸顺序和活性中心结构。Objective:To clone gene sequence of defibrase from venom gland of Gloidius ussuriensis and gett gene product of defibrase in the future. Methods:Total RNAs were extracted from the venom gland of the Gloydins Ussurisis snake.The thrombin like enzyme gene was amplified by RT PCR, cloned, and its nucleotide sequences had been determined.Results:The enzyme called defibrase cDNA encodes 702 nucleotides, namely 234 amino acids. Based on the homology, the catalytic residues and disulfide bridges of defibrase were deduced as followings: catalytic residues, His 41 ,Asp 86 , Ser 180 and six disulfide bridges. Conclusion:cDNA sequence of defibrase was proved, amino acid sequence and structure of active center were deduced.

关 键 词：乌苏里蝮蛇 降纤酶 克隆 序列分析 

分 类 号：Q785[生物学—分子生物学] R284.2[医药卫生—中药学]