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| 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 | |
| 引用本文： | 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) |
| 作者姓名： | 郭红敏 谷新晰 张玉兰 田洪涛 郭兴华 罗云波 |
| 作者单位： | 1. 河北农业大学食品科技学院,河北保定,071001 2. 中国科学院徽生物研究所,北京,100080 3. 中国农业大学食品科技与营养工程学院,北京,100083 |
| 基金项目： | 国家高技术研究发展计划(863计划)项目 |
| 摘 要： | 以典型的乳酸菌组成型表达质粒pMG36e为载体,德式乳杆菌保加利亚亚种为受体.采用电转化方法研究受体菌株的生长状态与细胞壁处理方式、质粒浓度、电转化参数、复苏培养基组成与复苏培养时间、受体菌株的限制修饰作用对电转化效率的影响,探讨电转化的最适条件,以提高电转化效率.结果表明:选择含2.5%甘氨酸的SMRS培养基培养受体细胞至对数中期,收获制成感受态细胞,在电场强度12 kV/cm、电阻200Ω、电容25μF条件下电击转化,以含20 mmol/L MgCl2、CaCl2的SMRS培养基复苏培养2 h,涂板筛选得到较高的电转化效率.特别是采用受体菌株修饰的质粒进行电转化,转化效率提高30倍以上,达到6.3×104cfu/μg DNA.本研究结果为进一步构建乳酸菌组成型高效表达系统和食品级基因工程乳酸菌株提供试验依据. |
| 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 |
Research of Electrotransformation Processing of Lactobacilli Delbrueckii Subsp.Bulgaricus with the Constitutive Expression Vector pMG36e |
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| Guo Hongmin,Gu Xinxi,Zhang Yulan,Tian Hongtao,Guo Xinghua,Luo Yunbo. Research of Electrotransformation Processing of Lactobacilli Delbrueckii Subsp.Bulgaricus with the Constitutive Expression Vector pMG36e[J]. Journal of Chinese Institute of Food Science and Technology, 2011, 11(2) | |
| Authors: | Guo Hongmin Gu Xinxi Zhang Yulan Tian Hongtao Guo Xinghua Luo Yunbo |
| Affiliation: | Guo Hongmin1 Gu Xinxi1 Zhang Yulan1 Tian Hongtao1 Guo Xinghua2 Luo Yunbo3 (1Food Science and Technology College,Agricultural University of Hebei,Baoding 071001,Hebei 2Institute of Microbiology,Academia Sinica,Beijing 100080 3College of Food Science and Nutritional Engineering,China Agricultural University,Beijing 100083) |
| 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 electroporation.Factors include wall treatment,plasmid concentration,electrotransformation 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 ... |
| Keywords: | Lactobacillus delbrueckii subsp.bulgaricus constitutive expression vector plasmid pMG36e electrotransformation electrotransformation efficiency |
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