摘要
来源于嗜热脂肪芽胞杆菌Bacillusstearothermophilus的耐热β 半乳糖苷酶基因bgaB被克隆到大肠杆菌-枯草芽胞杆菌穿梭质粒pMA5中,然后将外源基因及其表达调控序列亚克隆到枯草芽胞杆菌整合载体pSAS144中,转化Bacillussubtilis受体菌BD170,在5μg/mL的氯霉素抗性平板上筛选抗性转化子.经摇瓶发酵后,得到耐热乳糖酶的比酶活为0.32U/mg,是出发菌株比酶活的2倍.
The thermostable beta-galactosidase gene bgaB from Bacillus stearothermophilus was cloned into E.coli- B.subtilis. shuttle vector pMA5. Then bgaB gene and its regulatory sequence were subcloned to B.subtilis. integrative vector pSAS144 and formed a new integrative vector pSAS144-bgaB.This vector was then transformed to a chloramphenicol. Sensitive host B.subtilis BD170 and the transformants were received on LB agar medium with chloramphenicol to a final concentration of 5 μg/mL.The integrative strain showed thermostable -galactosidase activity of 0.32 U/mg, twice of that from original strain Bacillus stearothermophilus.
出处
《无锡轻工大学学报（食品与生物技术）》
CSCD
北大核心
2003年第6期1-4,14,共5页
Journal of Wuxi University of Light Industry
关键词
耐热β-半乳糖苷酶
枯草芽孢杆菌
基因表达
整合
thermostable beta-galactosidase
Bacillus subtilis
integration and expression
