大肠杆菌aroG基因在枯草杆菌中的分泌表达

为了对大肠杆菌中由ａｒｏＧ基因编码的３－脱氧－Ｄ－阿拉伯庚酮糖酸－７－磷酸合成酶（ＤＡＨＰ合成酶）进行结构与功能的深入研究，尝试了ａｒｏＧ基因在枯草杆菌中的表达。表达载体以ｐＵＢ１１０为骨架，采用了枯草杆菌热激蛋白ｇｒｏＥＳＬ基因的启动子，碱性蛋白酶ｓｕｂｔｉｌｉｓｉｎ基因的信号肽和Ｎ－乙酰胞壁酸－Ｌ－丙氨酸酰胺酶ｃｗｌＨＢ基因的转录终止子，将ａｒｏＧ基因在枯草杆菌ＷＢ６００宿主菌中进行了分泌表达

Secretory Expression of Escherichia coli aro G Gene in Bacillus subtilis

In Escherichia coli , the key enzyme for biosynthesis of aromatic amino acids, 3 deoxy D arabino heptulosonate 7 phosphate (DAHP) synthase are encoded by three genes, aro G, aro H and aro F. This study showed the expression of E.coli aro G gene in Bacillus subtilis as secreted proteins to benefit the purification and crystallization of the enzyme for structure and function study. The aro G gene was subcloned into expression vector pUB110, with gro ESL promoter, subtilisin signal peptide coding DNA upstream and cwl B terminator downstream. The band corresponding to aro G gene product was shown on SDS polyacrylamide gel electrophoresis, and the DAHP syntase acivity was detected in the culture medium.