Yang Qing, Hu Xue-Jun, Xu Xiao-Ming, An Li-Jia, Yuan Xiao-Dong, Su Zhi-Guo
Bioengineering Institute, Dalian University of Technology, Dalian 116012, China.
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai). 2002 Jan;34(1):6-10.
Total RNAs were extracted from the venom gland of the snake Gloydius ussuriensis. The cDNA of gussurobin, a thrombin-like enzyme from Gloydius ussuriensis, was cloned and amplified by RT-PCR. Assay of the nucleotide sequence of the cDNA allowed postulation of the complete amino acid sequence for Gloydius ussuriensis, Chinese Viperdae. Its amino acid sequence exhibits significant homology with that of other snake thrombin-like enzymes. The cDNA of gussurobin was inserted into the vector pPIC 9K and expressed successfully in Pichia pastoris, strain GS115. The recombinant gussurobin was separated and purified from 500 ml culture and showed one band on SDS-PAGE.
从乌苏里蝮蛇的毒腺中提取总RNA。通过RT-PCR克隆并扩增了乌苏里蝮蛇一种类凝血酶（gussurobin）的cDNA。对该cDNA核苷酸序列的分析推测出了乌苏里蝮蛇（中国蝰科）完整的氨基酸序列。其氨基酸序列与其他蛇类类凝血酶的氨基酸序列具有显著同源性。将gussurobin的cDNA插入载体pPIC 9K，并在毕赤酵母GS115菌株中成功表达。从500毫升培养物中分离并纯化出重组gussurobin，其在SDS-PAGE上显示出一条带。
