摘要
目的:研究山梨糖还原酶基因sr(B932_3022)在氧化葡糖杆菌1.637生长代谢中的作用。方法:构建sr基因同源敲除质粒p GX-srup-Gm-down,PCR扩增得到打靶片段,电击转化导入氧化葡糖杆菌1.637,通过庆大霉素抗性和PCR筛选sr基因重组敲除菌,分别以山梨醇和山梨糖为碳源考察野生菌与敲除菌生长和山梨糖代谢的差异。结果:抗性和PCR验证结果显示敲除了氧化葡糖杆菌1.637的sr基因;与野生菌相比,敲除菌生长出现滞后,以山梨醇为碳源时敲除菌山梨糖积累增多,而以山梨糖为碳源时山梨糖消耗减少。结论:氧化葡糖杆菌1.637的sr基因敲除影响菌株前期生长与山梨糖代谢。
Objective： To study the function of sorbose reductase gene sr（B932_3022） from Gluconobacter oxydans 1.637. Methods： Plasmid p GX-srup-Gm-down was constructed and homologous fragment was transfected intocompetent cell of G.oxydans 1.637 by electroporation. The deletion mutant of sr（1.637Δsr） was verified by gentami-cin resistant and PCR method. The differences of biological characteristics such as growth rate and the level ofsorbose metabolism between the 1.637Δsr and the wild strain were investigated. Results： The sr gene was success-fully knocked out. Compared with the wild strain 1.637, the mutant strain showed a lag phrase, increased accumu-lation of sorbose in YBH media and decreased consumption of sorbose in TBHT media. Conclusion： The deletionof sr gene may influence the growth rate and the metabolism of sorbose of G.oxydans 1.637.
出处
《生物技术通讯》
CAS
2016年第2期192-195,共4页
Letters in Biotechnology
