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| 乳酸杆菌组成型启动子探测载体pgateway-612的构建 | |
| 引用本文： | 陈家锃,崔红玉,田志军,葛俊伟,安同庆,彭金美,李一经.乳酸杆菌组成型启动子探测载体pgateway-612的构建[J].中国预防兽医学报,2011,33(11). |
| 作者姓名： | 陈家锃 崔红玉 田志军 葛俊伟 安同庆 彭金美 李一经 |
| 作者单位： | 1. 东北农业大学动物医学院,黑龙江哈尔滨150030;中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,黑龙江哈尔滨150001 2. 东北农业大学动物医学院,黑龙江哈尔滨,150030 3. 中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,黑龙江哈尔滨,150001 |
| 基金项目： | 国家十一五支撑计划项目 |
| 摘 要： | 为筛选适宜乳酸菌(LAB)宿主表达载体的启动子,本研究以大肠杆菌- LAB穿梭载体pPG612为基本骨架,利用GatewayTM克隆技术操作构建了LAB组成型启动子探测载体,并利用该载体筛选了短乳杆菌的S层启动子.我们利用底物X-gluC与报告基因gusA编码的蛋白酶之间的特异性显色反应筛选阳性菌落,超声破碎后的菌体蛋白经SDS-PAGE和westem blot分析表明该启动子在宿主菌中具有组成型启动子活性.本研究为进一步利用表达载体及筛选其他LAB启动子奠定基础. |
| 关 键 词： | 乳酸菌 启动子探测载体 Gateway克隆技术 X-gluC |
Construction of a constitutive promoter probing vector in Lactobacillus |
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| CHEN Jia-zeng,CUI Hong-yu,TIAN Zhi-jun,GE Jun-wei,AN Tong-qin,PENG Jin-mei,LI Yi-jing.Construction of a constitutive promoter probing vector in Lactobacillus[J].Chinese Journal of Preventive Veterinary Medicine,2011,33(11). | |
| Authors: | CHEN Jia-zeng CUI Hong-yu TIAN Zhi-jun GE Jun-wei AN Tong-qin PENG Jin-mei LI Yi-jing |
| Institution: | CHEN Jia-zeng1,2,CUI Hong-yu1,TIAN Zhi-jun2,GE Jun-wei1,AN Tong-qing2,PENG Jin-mei2,LI Yi-jing1(1.College of Veterinary Medicine,Northeast Agricultural University,Harbin 150030,China,2.State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150001,China) |
| Abstract: | pPG612 is a shuttle vector suitiable for replication in both E.coli and Lactobacillus.In the current study,we replaced the promoter of pPG612 vector with S-layer protein promoter of Lactobaccillus brevis by using the GatewayTM cloning technology.The resultant expression vector was used for detection of expressing target gene in Lactobacillus.Positive clones were screened by using X-gluC,which was able to specifically enzymolied substance into blue color by the reporter protein Gus.We further confirmed that ... |
| Keywords: | Lactobacillus brevis promoter probing vector gateway clone X-gluC |
| 本文献已被 CNKI 万方数据 等数据库收录！ | |