【摘要】 目的 :构建人腺体激肽释放酶 (h K2 )基因的酵母表达载体。方法 :利用 RT- PCR法从人正常前列腺组织中扩增出 h K2基因 ,先将其克隆到 p GEM- T载体 ,进行序列测定和分析 ,然后将 h K2基因亚克隆至酵母p PICZαC表达载体上 ,进行鉴定。结果 :获得一个核苷酸长度为 738bp的基因 ,同源比较结果表明 ,与 Gen Bank(NCBI:AF18874 6 )公布的 h K2的基因序列有 99%的同源性 ,序列分析表明有一个氨基酸出现变异。酶切表明 h K2基因正确插入酵母表达载体 p PICZαC。结论 :成功构建出 h K2基因的酵母表达载体 p PICZαCh K2。更多还原

【Abstract】 Objective To construct yeast expression vector which can express human glandular kallikrein highly. Methods The hK2 gene was amplified by RT-PCR from human normal prostate tissue. The amplified DNA fragment was cloned into pGEM-T vector for sequence analysis. The hK2 gene was then subcloned into yeast expression vector pPICZαC. Results A gene of 738bp was obtained and showed a 99% homology with hK2 gene sequence published in GenBank(NCBI: AF188746).Sequence analysis showed that only one amino acid was different.hK2 gene was inserted into yeast expression vector pPICZαC correctly. Conclusion The hK2 yeast expression vector pPICZαChK2 is constructed successfully in the study.更多还原