【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。更多还原

【Abstract】 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.更多还原