枯草芽孢杆菌
转化效率
电穿孔
转化(遗传学)
质粒
生物
穿梭机载体
杆菌科
DNA
山梨醇
海藻糖
微生物学
细菌
基因
遗传学
生物化学
载体(分子生物学)
重组DNA
农杆菌
作者
Yingjian Lu,C. Zhang,Fengting Lv,Xiaomei Bie,Zhaoxin Lu
标识
DOI:10.1111/j.1472-765x.2012.03249.x
摘要
Aims: To optimize the transformation conditions and improve the transformation efficiency of Bacillus subtilis WB800 and DB104. Methods and Results: Trehalose, which could decrease the damage of electric shock to the cells, was added to the electroporation medium containing sorbitol and mannitol. The factors affecting the transformation efficiency, such as the growth phase of bacteria, cell concentration, electric field strength and plasmid variety, were examined and improved. The new method increased the transformation efficiency of B. subtilis by nearly 100‐fold compared with the conventional one. Conclusions: With the optimized method, the transformation efficiency came up to 3·64 × 105 transformants μg−1 DNA for WB800, and 2·10 × 105 transformants μg−1 DNA for DB104. Significance and Impact of the Study: This improvement in transformation efficiency will be largely attributed to the research of expression of exogenous genes in B. subtilis, gene library construction for directed evolution and transformation of wild‐type B. subtilis strains.
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