摘要
黑曲霉糖化酶基因cDNA按正确的读码框架克隆到广宿主表达载体pBBR1MCS-2,构建出含黑曲霉糖化酶基因的表达载体pBBR1MCS-2GA,用三亲和接合转化法将重组质粒导入2-酮基-D-葡萄糖酸高产菌株荧光假单胞菌AR4。得到重组菌ARW4,经SDS-PAGE凝胶电泳和Westernblot分析显示,黑曲霉糖化酶基因在ARW4中得到表达,表达产物主要以不溶性包涵体形式存在。
Aspergillus niger glucoamylase gene cDNA was subcloned into broad host expression vector pBBR1MCS-2 with correct reading frame to obtain recombinant vector pBBR1MCS-2GA. Then it was transformed into Pseudomonasfluorescens AR4 which was a high yield strain of 2-keto-D-gluconic acid, and obtained recombinant bacteria ARW4. SDS-PAGE gel electrophoresis and Western blot analysis showed that the recombinant glucoamylase protein had expressed effectively in ARW4, and existed in the form of inclusion which was inactive.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2006年第11期199-202,共4页
Food Science
关键词
黑曲霉
糖化酶基因
荧光假单胞菌
原核表达
Aspergillus niger
glucoamylase gene
Pseudomonas fluorescens
prokaryotic expression
