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乳酸乳球菌乳脂亚种MG1363中盐诱导启动子的克隆
引用本文:余红仙,许文涛,程国灵,戴蕴青,黄昆仑,田洪涛,罗云波.乳酸乳球菌乳脂亚种MG1363中盐诱导启动子的克隆[J].食品工业科技,2012,33(11):187-190.
作者姓名:余红仙  许文涛  程国灵  戴蕴青  黄昆仑  田洪涛  罗云波
作者单位:1. 中国农业大学食品科学与营养工程学院,北京,100083
2. 中国农业大学食品科学与营养工程学院,北京100083/农业部转基因生物使用安全检验监督测试中心,北京100083
3. 农业部转基因生物使用安全检验监督测试中心,北京,100083
4. 河北农业大学食品科技学院,河北保定,071001
基金项目:农业部“948”项目(2007-Z8);国家自然基金(30800770);转基因生物重大专项(2008ZX08012-001)
摘    要:为了从乳酸菌中筛选和克隆启动子,实验利用缺失T7启动子的质粒载体PRSET/LacZ直接在大肠杆菌(E.coli)DH5α中分离乳酸乳球菌乳脂亚种(Lactococcus lactis subsp.cremoris)MG1363的基因启动子片段,获得了10多个具有抗氨苄和盐诱导出蓝斑的重组子。反复筛选并对其中一个抗性最高的重组子PRSET-osm进行序列测定和同源性分析发现,所克隆的基因启动子片段来自乳酸乳球菌乳脂亚种MG1363的基因组,并具有原核启动子的保守序列(Pribnow框和Sextama框)。对启动子osm进行进一步序列分析和鉴定发现,其在大肠杆菌BL21中启动LacZ基因的表达,确定osm为盐诱导启动子。

关 键 词:乳酸乳球菌乳脂亚种MG1363  大肠杆菌  基因启动子

Cloning of promoters from Lactococcus lactis subsp.cremoris MG1363
YU Hong-xian,XU Wen-tao,CHENG Guo-ling,DAI Yun-qing,HUANG Kun-lun,TIAN Hong-tao,LUO Yun-bo.Cloning of promoters from Lactococcus lactis subsp.cremoris MG1363[J].Science and Technology of Food Industry,2012,33(11):187-190.
Authors:YU Hong-xian  XU Wen-tao  CHENG Guo-ling  DAI Yun-qing  HUANG Kun-lun  TIAN Hong-tao  LUO Yun-bo
Affiliation:1,2,*(1.College of Food Science and Nutritional Engineering,China Agricultural University,Beijing 100083,China; 2.The Supervision,Inspection & Testing Center of Genetically Modified Orginisms Food Safety,Ministry of Agriculture, Beijing 100083,China; 3.College of Food Science and Technology,Hebei Agricultural University,Baoding 071001,China)
Abstract:For the purpose of screening and cloning promoters from Lactococcus lactis cremoris MG1363,DNA fragments were separated by using the T7 promoter-less vector PRSET/LacZ in E.coli DH5α.More than 10 fragments were obtained,which were identified based on their ability to confer resistance against Ampicillin and induced by high press.After sequencing and homeology blast,a recon was found(PRSET-gro).It was from Lactococcus lactis cremoris MG1363 and has the identical consensus region Pribnow box and Sextama box.After sequences analysis and identification of osm putative promoter,we found that it could endow expression of LacZ in E.coli BL21,moreover,the osm promoter was intended to be osmotic pressure inducible promoter.
Keywords:Lactococcus lactis subsp  cremoris MG1363  E  coli  promoter
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