摘要
通过PCR扩增了肺炎克雷伯氏菌(Klebsiella pneumoniae)和大肠杆菌(Escherichia coli)各自的葡萄糖脱氢酶基因KPgdh和ECgdh,构建了表达载体p ET-28a-KPgdh和p ET-28a-ECgdh,转化大肠杆菌E.coli BL21,获得了重组菌BL21(p ET-28a-KPgdh)和BL21(p ET-28a-ECgdh)。经IPTG诱导,聚丙烯酰胺凝胶电泳显示重组菌实现了葡萄糖脱氢酶的高效表达,且葡萄糖脱氢酶活性分别是空质粒对照菌E.coli BL21(p ET-28a)的8.5倍和12倍。重组菌的生长速度快于空质粒对照菌E.coli BL21(p ET-28a),并与原代菌E.coli BL21持平。如果扣除质粒复制造成的代谢负荷,过表达葡萄糖脱氢酶促进了大肠杆菌的生长。
The genes encoding glucose dehydrogenase（gdh） from Klebsiella pneumoniae and Escherichia coli were amplified by polymerase chain reaction（PCR）. Subsequently,the corresponding expression vectors p ET-28a-KPgdh and p ET-28a-ECgdh were constructed and cloned into E. coli BL21,resulting in two recombinant strains E. coli（p ET-28a-KPgdh） and E. coli（p ET-28a-ECgdh）. SDS-PAGE analysis showed the high level expression of both KPgdh and ECgdh upon IPTG induction. Compared with the control strain E. coli（p ET-28a）which harbored an empty vector,the recombinant strains E. coli（p ET-28a-KPgdh） and E. coli（p ET-28a-ECgdh） consumed more glucose,and the gdh activity toward glucose increased by 8.5-and 12-fold,respectively. If plasmid burden was not counted,overall these results indicated that gdh overexpression facilitated cell growth.
出处
《食品工业科技》
CAS
CSCD
北大核心
2015年第22期189-192,197,共5页
Science and Technology of Food Industry
基金
国家自然科学基金(21076013
21276014)
关键词
大肠杆菌
肺炎克雷伯氏菌
葡萄糖脱氢酶
超表达
细胞生长
Escherichia coli
Klebsiella pneumoniae
glucose dehydrogenase
overexpression
cell growth
