α-乙酰乳酸脱羧酶在毕赤酵母中的分泌表达  

Secreted Expresson α-Acetolactate Decarboxylase in Pichia pastoris

作　　者：李联正[1] 张惟材[1] 汪建华[1] 何维明[2] 

机构地区：[1]军事医学科学院生物工程研究所,北京100071 [2]西北农林科技大学动物科技学院,陕西杨凌712100

出　　处：《生物技术通讯》2005年第6期621-623,共3页Letters in Biotechnology

摘　　要：根据已知的α-乙酰乳酸脱羧酶(ALDC)基因序列,通过PCR从枯草杆菌168基因组上扩增得到编码ALDC的结构基因。将该基因克隆到大肠杆菌-毕赤酵母穿梭载体pPIC9中,构建了重组质粒pPIC9-ALDC,电击转化毕赤酵母GS115。在甲醇诱导下,毕赤酵母分泌表达了ALDC。SDS-PAGE分析可见到Mr约62000的表达条带,经测定,表达产物活力为0.03U/mL。α-acetolactate decarboxylase（ALDC） gene was generated from genomic DNA of Bacillus subtilis 168 by PCR, then cloned into an E, coli-yeast shuttle plasmid pPIC9. A recombinant plasmid pPIC9-ALDC was thus constructed and transformed into competent Pichia pastoris strain GSI15. It is shown that the recombinant, GSII5/pPIC9-ALDC, secreted ALDC under induction with methanol, A band of 62 kDa was detected under SDS-PAGE assay. The biological activity of the recombinant is determined to be 0.03 U/mL.

关 键 词：Α-乙酰乳酸脱羧酶 枯草芽孢杆菌 分泌表达 活性测定 

分 类 号：Q785[生物学—分子生物学] Q786