| 利用E.coli-App穿梭载体构建温控双基因裂解载体pMC-WK |
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| 引用本文： | 孙颖,金天明,冯立文,袁洁.利用E.coli-App穿梭载体构建温控双基因裂解载体pMC-WK[J].中国兽医学报,2011,31(9). |
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| 作者姓名： | 孙颖 金天明 冯立文 袁洁 |
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| 作者单位： | 广东省农业科学院兽医研究所,广东广州,510640 |
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| 基金项目： | 天津市高等学校科技发展基金计划项目(20060720); 国家自然科学基金资助项目(31072109) |
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| 摘 要： | 采用PCR方法从噬菌体PhiX174基因组中扩增出裂解蛋白E基因,从金黄色葡萄球菌基因组中扩增出核酸酶A基因（SN）,通过15个柔性氨基酸linker将双基因串联,插入PBV220表达载体,构建温控双基因裂解系统（DLS）。PCR扩增DLS,将其与E.coli-App穿梭载体pMC-Express相连,构建重组穿梭载体pMC-WK。裂解试验表明,E-15aalinker-SN裂解较E裂解迅速而彻底,菌体裂解率达到99.999 4%;经PCR扩增出的DLS具有裂解活性。本试验成功构建了温控双基因裂解载体pMC-WK,为App菌影疫苗的研究奠定了基础。
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| 关 键 词： | 裂解蛋白E 核酸酶A App菌影 穿梭载体 |
Construction of E.coli-App temperature controlled double gene cracking vect or pMC-WK |
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| SUN Ying,JIN Tian-ming,FENG Li-wen,YUAN Jie.Construction of E.coli-App temperature controlled double gene cracking vect or pMC-WK[J].Chinese Journal of Veterinary Science,2011,31(9). |
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| Authors: | SUN Ying JIN Tian-ming FENG Li-wen YUAN Jie |
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| Institution: | SUN Ying1,JIN Tian-ming1,FENG Li-wen2,YUAN Jie3 (1.Animal Science Department,Tianjin Agricultural College,Tianjin 300 384,China,2.College of Aminal Science and Veterinary Medicine,Jilin University Changchun 130062,3.Institute of Guangdong Veterinary Science,Guangzhou 510640,China) |
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| Abstract: | Using the PCR method to clone lysis protein E gene from the genome of phage PhiX174 and nuclease A gene(SN) from the genome of Staphylococcus aureus. Link the two genes with 15 amino acid flexible linker to get double-gene tandem, then insert it into PBV220 plasmid.Amplifing DLS through PCR to construct pMC-W K on the basis of E.coli-App shuttle vector pMC-Express.Lysis experiments s how th at E-15aalinker-SN system has better cracking efficiency than E gene system.Th e lysis rate can be 99.999 4% and DLS g... |
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| Keywords: | lysis protein E nuclease A App bacterial ghost shuttle vector |
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