| 大肠杆菌“菌影”的制备 |
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| 引用本文： | 彭玮,王聃,刘思国,常月红,王春来,刘慧芳,司薇,赫明雷,杜艳芬. 大肠杆菌“菌影”的制备[J]. 东北农业大学学报, 2009, 40(6) |
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| 作者姓名： | 彭玮 王聃 刘思国 常月红 王春来 刘慧芳 司薇 赫明雷 杜艳芬 |
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| 作者单位： | 1. 东北农业大学动物医学学院,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动物细菌病研究室,哈尔滨,150001
2. 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动物细菌病研究室,哈尔滨,150001 |
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| 基金项目： | 国家高技术研究发展计划 |
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| 摘 要： | 通过PCR扩增噬菌体PhiX174的裂解基因E,将该基因连接到含有PR/cI启动阻遏系统的pBV220载体中,从而使裂解基因E和启动阻遏系统λPL/PR-cI857串联成为温度敏感的裂解盒,构建溶菌质粒pBV-E。采用CaCl2法将其转入大肠杆菌BL21(DE3),含有裂解质粒的大肠杆菌在28℃条件下生长到对数生长期,然后升温到42℃诱导其溶解,制成了大肠杆菌菌影。电镜观察发现菌影形态完整,内容物全部被释放到胞外。
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| 关 键 词： | 大肠杆菌 菌影 裂解基因E |
Generation of Escherichia coli ghosts |
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| PENG Wei,WANG Dan,LIU Siguo,CHANG Yuehong,WANG Chunlai,LIU Huifang,SI Wei,HE Minglei,DU Yanfen. Generation of Escherichia coli ghosts[J]. Journal of Northeast Agricultural University, 2009, 40(6) |
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| Authors: | PENG Wei WANG Dan LIU Siguo CHANG Yuehong WANG Chunlai LIU Huifang SI Wei HE Minglei DU Yanfen |
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| Affiliation: | 1. College of Veterinary Medicine;Northeast Agricultural University;Harbin 150030;China;2. National Key Laboratory of Veterinary Biotechnology;Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences;Harbin 150001;China |
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| Abstract: | In the present study, lytic gene E from PhiX174 was amplified by PCR and inserted into pBV220 vector which included the λPL/PR-cI857 regulatory system. Lytic gene E was connected in series with the λPL/PR- cI857 regulatory system to form the temperature-sensitive lysis case that was in constructed bacteriolysis plasmid pBV-E. The plasmid pBV-E was transformed into E. coli BL21(DE3) by calcium chloride method. E. coli with the gene E was grown in a waterbath shaker at 28 ℃ under aerobic conditions until mid log-phase. Then the culture was transferred to 42 ℃ water-bath shaker. Finnally, E.coli ghosts were produced, which were in intact forms under transmission electron microscope, and their contents were released to extra cellular region. This experiment laid a foundation of study on bacterial ghost as a novel vaccine and form of adjuvant. |
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| Keywords: | E. coli ghost lytic gene E |
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