摘要
利用鸟枪法构建了供体菌的基因组文库，通过VP反应显色法进行筛选，从约7000个转化子中选出携带ALDC基因的重组质粒（pBG4～pBG5）．绘制了pBG4插入片段的限制酶图谱．Southern杂交证明该外源片段来源于供体菌。酶活性测定结果表明ALDC基因在大肠杆菌中获得了表达，为构建带有ALDC的啤酒酵母工程菌奠定了基础．
Bacillus subtilis BD-5 genomic library has been constructed using plasmid pUC18 in Escherichia colt. The hybrid plasmids (pBG3~5) which contained the gene encoding α- acetolactate decaroboxylase (ALDC) have been isolated from thes library by Voges-Proskauer tests. Southern hybridizantion showed that the inserted fragment of pBG4 was from the BD-5.The restrichon map of the insert fragment of pBG4 has been drawn. The ALDC activety was expressed in the Escherichia coli. This work laid a foundation to constuct the genetically-modified brewing yeasts with ALDC gene.
出处
《微生物学通报》
CAS
CSCD
北大核心
1998年第6期336-339,共4页
Microbiology China
基金
国家自然科学基金
关键词
枯草芽孢杆菌
ALDC基因
克隆
表达
Bacillus subtilis, α-Acetolactate decaroboxylase gene, Cloning and Expression
