破菌时可自动降解宿主核酸大肠杆菌BL21(DE3)lpxM突变株构建.pdfVIP

生 物 工 程 学 报 Chin J Biotech 2008, January 25; 24(1): 46-52 Chinese Journal of Biotechnology ISSN 1000-3061 cjb@ ©2008 Institute of Microbiology, CAS CSM, All rights reserved. 研究报告 BL21(DE3) lpxM 1, 2 1 1 3 1 张 君 , 方宏清 , 戴红梅 , 谢达平 , 陈惠鹏 1 , 100071 2 410004 3 , 410128 : 研究利用 Red 同源重组技术对常用大肠杆菌表达宿主菌 BL21(DE3)进行改良, 构建破菌时可自动降解宿主核 酸的大肠杆菌表达宿主菌, 该菌株可望有助于解决因破菌时宿主菌染色体核酸释放给后续纯化重组蛋白工作带来的困 难。将 N 端连有 OmpA 的信号肽的 S. aureus nucleaseB(nucB)表达框整合至 E. coli BL21(DE3) 的 lpxM 位点, 改造后菌株 (称为 BLN)经诱导能表达 nucB 、并分泌至周质空间, 这样可使宿主核酸免受该酶“毒性”影响, 菌体裂解后, nucB 释放, 能自动降解宿主核酸。BLN 菌体生长状态以及表达外源重组蛋白的能力与出发菌基本一致。 : E.coli BL21(DE3), Red 同源重组系统, lpxM, 金黄色葡萄球菌核酸酶 Constructing Modified Protein-producing Escherichia coli Capable of Autohydrolysing Host Nucleic Acid During Cell Lysis 1, 2 1 1 3 1 Jun Zhang , Hongqing Fang , Hongmei Dai , Daping Xie , and Huipeng Chen 1 Institute of Biotechnology , Academy of Military Medical Sciences , Beijing 100071, China 2 Hunan Vocational College of Science and Technology, Changsha 410004, China 3 School of Biological Sciences, Hunan Agricultural University, Changsha 410128, China Abstract: We used Red recombination system to construct a modified E. coli protein-producing strain capable of autohydrolysing host nucleic acid. E. coli BL21(DE3), a common protein-producing strain, was used as starting