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作 者:高彤 李明桂 梁水明 范丽佳 王长丽 Gao Tong;Li Minggui;Liang Shuiming;Fan Lijia;Wang Changli(Youjiang Medical College for nationalities,School of clinical medicine,GuangxiBaise 533000;Youjiang Medical College for nationalities,Medical laboratory College,GuangxiBaise 533000)
机构地区:[1]右江民族医学院临床医学院,广西百色533000 [2]右江民族医学院基础医学院,广西百色533000
出 处:《科技风》2023年第12期162-165,共4页
基 金:广西高校中青年教师基础能力提升项目(2020KY13007);自治区教育厅2021年和2022年创新创业训练计划项目(S202110599090、S202210599042);右江民族医学院2021年度校级科研课题(yy2021sk063);2021年右江民族医学院“新启航”青年骨干项目。
摘 要:目的:构建、筛选并鉴定出重组GFP唾液乳杆菌CCFM6105。方法:利用GFP作为报告基因,从携带gWiz-GFP质粒的大肠杆菌中提取质粒,将gWiz-GFP质粒电转化至唾液乳杆菌CCFM6105中,构建GFP唾液乳杆菌。结果:PCR产物电泳结果显示约在750bp处出现明亮条带,PCR反应成功;在1800V电压下的菌落荧光暗沉;在2400V电压下的菌落荧光致密明亮;在2200V、2600V电压下菌落荧光偏暗;各代重组菌的GFP表达良好且无明显减弱。结论:本实验成功构建GFP唾液乳杆菌CCFM6105且GFP在唾液乳杆菌在稳定遗传中得到了稳定的表达。Objective:To construct,screen and identify recombinant GFP Lactobacillus salivarius CCFM6105.Methods:using GFP as the reporter gene,the plasmid was extracted from E.coli carrying gWiz GFP plasmid,and the gWiz GFP plasmid was electrotransformed into L.salivarius CCFM6105 to construct L.salivarius GFP.Results:The electrophoresis results of PCR products showed that a bright band appeared at about 750bp,and the PCR reaction was successful;Colony fluorescence darkening at 1800V voltage;The colony fluorescence was dense and bright at 2400V voltage;The colony fluorescence was dark at 2200V and 2600V;The expression of GFP in the recombinant strains of each generation was good without obvious weakening.Conclusion:In this experiment,GFP in L.salivarius CCFM6105 was successfully constructed and GFP in L.salivarius was stably expressed in the stable genetic.
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