中国食品学报2011，Vol.11Issue(2)：8-16,9.
利用组成型表达载体pMG36e电转化德式乳杆菌保加利亚亚种的研究
Research of Electrotransformation Processing of Lactobacilli Delbrueckii Subsp.Bulgaricus with the Constitutive Expression Vector pMG36e
摘要
Abstract
Using the constitutive expression plasmid pMG36e of the lactic acid bacteria as a vector, and the Lactobacilli delbrueckii subsp bulgaricus as a host,the different factors on the impact of electroporation efficiency was studied by the method of electroporafion. Factors include wall treatment, plasmid concentration, electrotransformafion parameters,composition of the recovery medium, recovery culture time and the restriction modification of host. Results showed that the optimal values for concentration of glycine and of sucrose and the pulse strength、risistance are 2.5％, 0.5 mol/L and 12 kV/cm, 200 Ω, respectively. Best electrotransformation efficiency was attained after incubation in 20 mmol/L MgCl2、 CaCl2 containing SMRS broth for another 2 h,combined with the antibiotics in the selective plates. When the receptormodified strains of plasmid was used to electrotransformation, the transformation efficiency has increased by 30 times,reached 6.3×104 cfu/μg DNA. The research of this paper provided foundation for further study of establish high-efficiency expression system of lactic acid bacteria and the development of genetic engineering technology.关键词
德式乳杆菌保加利亚亚种/组成型表达载体/质粒pMG36e/电转化/电转化效率Key words
Lactobacillus delbrueckii subsp bulgaricus/ constitutive expression vector/ plasmid pMG36e/ electrotransformation/ electrotransformation efficiency引用本文复制引用
郭红敏,谷新晰,张玉兰,田洪涛,郭兴华,罗云波..利用组成型表达载体pMG36e电转化德式乳杆菌保加利亚亚种的研究[J].中国食品学报,2011,11(2):8-16,9.基金项目
国家高技术研究发展计划(863计划)项目(No.2006AA10Z317) （863计划）
