| 基于CRISPR/Cas9系统对干酪乳杆菌进行红色荧光蛋白标记 |
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| 引用本文： | 张莹,宋馨,夏永军,艾连中,王光强.基于CRISPR/Cas9系统对干酪乳杆菌进行红色荧光蛋白标记[J].工业微生物,2019,49(1):14-20. |
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| 作者姓名： | 张莹 宋馨 夏永军 艾连中 王光强 |
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| 作者单位： | 上海理工大学 医疗器械与食品学院,上海,200093;上海理工大学 医疗器械与食品学院,上海,200093;上海理工大学 医疗器械与食品学院,上海,200093;上海理工大学 医疗器械与食品学院,上海,200093;上海理工大学 医疗器械与食品学院,上海,200093 |
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| 摘 要： | 本实验利用CRISPR/Cas9系统对干酪乳杆菌(Lactobacillus casei) LC2W进行红色荧光蛋白(red fluorescent protein,RFP)标记,用于研究干酪乳杆菌在肠道内的分布和定植状况,评价其作为益生菌的功能。首先,基于本实验室已有的干酪乳杆菌CRISPR/Cas9编辑质粒pLCNICK-1628构建重组质粒pLCNICK-1628-RFP,电转入干酪乳杆菌LC2W感受态细胞中,使干酪乳杆菌基因组中的LC2W-1628基因被红色荧光蛋白基因替换,从而使干酪乳杆菌LC2W能表达出红色荧光蛋白。得到红色荧光标记的干酪乳杆菌LC2W突变株后,测定了其荧光强度-OD600标准曲线,发现RFP在干酪乳杆菌LC2W中能稳定表达。
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| 关 键 词： | 干酪乳杆菌 LC2W CRISPR/Cas9系统 红色荧光蛋白 |
Red fluorescent protein labeling of Lactobacillus casei based on CRISPR/Cas9 system |
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| ZHANG Ying,SONG Xin,XIA Yongjun,AI Lianzhong,WANG Guangqiang.Red fluorescent protein labeling of Lactobacillus casei based on CRISPR/Cas9 system[J].Industrial Microbiology,2019,49(1):14-20. |
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| Authors: | ZHANG Ying SONG Xin XIA Yongjun AI Lianzhong WANG Guangqiang |
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| Institution: | (School of Medical Instrument and Food Engineering,University of Shanghai for Science andTechnology,Shanghai 200093,China) |
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| Abstract: | In this study,CRISPR/Cas9 system was used to label Lactobacillus casei LC2W with red fluorescent protein(RFP)for investigating the distribution and colonization of Lactobacillus casei in intestinal tract and evaluating its function as probiotics.First,the plasmid pLCNICK-1628-RFP was constructed and was introduced into the receptor cells of LC2W.Then,the LC2W-1628 gene in genome of LC2W was deleted and the RFP gene was inserted into the genome of LC2W.Therefore,LC2W could express RFP.The results showed that RFP could be expressed stably in Lactobacillus casei LC2W by measuring the fluorescence intensity. |
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| Keywords: | Lactobacillus casei LC2W CRISPR/Cas9 system red fluorescent protein |
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