中国农业科学院农业信息研究所 中国农学会
Agricultural Information Institute of CAAS China Association of Agricultural Science Societies
En
Ch
中国农业科学院农业信息研究所 中国农学会 主办

AI检索

订阅

返回顶部
当前位置: 首页 > 文章 > 肠出血性大肠杆菌O157∶H7 toxB基因缺失株的构建 江苏农业学报 2012,28 (2) 337-343
Position: Home > Articles > Construction of enterohemorrhagic Escherichia coli O157∶H7 deficient in toxB gene Jiangsu Journal of Agricultural Sciences 2012,28 (2) 337-343

肠出血性大肠杆菌O157∶H7 toxB基因缺失株的构建

作  者：
张雪寒;何孔旺;赵攀登;栾晓婷;叶青;温立斌;倪艳秀;李彬;郭容利;王小敏;周俊明;俞正玉;茅爱华;吕立新
单  位：
江苏省农业科学院兽医研究所;农业部兽用生物制品工程技术重点实验室;国家兽用生物制品工程技术研究中心
关键词：
肠出血性大肠杆菌O157∶H7;toxB基因;缺失株;小鼠;黏附
摘  要：
为了明确肠出血性大肠杆菌(Enterohemorrhagic Escherichia coli)O157∶H7大质粒pO157编码的蛋白质ToxB与O157∶H7在肠道黏附、定殖的关系,构建含有toxB基因同源臂的重组自杀性质粒pMEG375-BN-Kan-BC,转化SM10宿主菌获得重组SM10(pMEG375-BN-Kan-BC)。将SM10(pMEG375-BN-Kan-BC)与O157∶H7进行混合培养,通过同源重组,获得杂交toxB基因缺失株。用体外接种的HEp-2细胞和体内感染链霉素处理的小鼠,明确缺失株黏附定殖能力。经PCR和Western blot鉴定,toxB基因被卡那霉素(Kan+)选择标记基因表达盒所代替。O157∶H7(△toxB)生长特性与亲本株相比无明显差异。O157∶H7(△toxB)与亲本株相比,对HEp-2细胞的黏附能力降低了2倍。口服感染小鼠后,O157∶H7(△toxB)第3 d粪便排菌量仅为3.1×105 CFU,持续排菌11 d;而亲本株排菌量高达4.03×107 CFU,持续排菌时间超过14 d。
译  名：
Construction of enterohemorrhagic Escherichia coli O157∶H7 deficient in toxB gene
作  者：
ZHANG Xue-han,HE Kong-wang,ZHAO Pan-deng,LUAN Xiao-ting,YE Qing,WEN Li-bin,NI Yan-xiu,LI-Bin,GUO Rong-li,WANG Xiao-min,ZHOU Jun-ming,YU Zheng-yu,MAO Ai-hua,Lü Li-xin(Institute of Veterinary Medicine,Jiangsu Academy of Agricultural Sciences/Key Laboratory of Engineering Research of Veterinary Bio-products,Ministry of Agriculture/National Center for Engineering Research of Veterinary Bio-products,Nanjing 210014,China)
关键词：
enterohemorrhagic Escherichia coli O157∶H7;toxB gene;knockout strain;mouse;adherence
摘  要：
In order to definite the correlation between ToxB,a large protein encoded by pO157 of enterohemorrhagic Escherichia coli O157∶H7,and intestinal adherence and colonization of O157∶H7,the recombinant suicide plasmid pMEG375-BN-Kan-BC was construted by cloning homologous arms of toxB upward and downward genes into suicide plasmid,and was transformed into host bacterium SM10 complement cells to obtain recombinant SM10(pMEG375-BN-Kan-BC).The SM10(pMEG375-BN-Kan-BC) was incubated with O157∶H7 to construct O157∶H7(△toxB) through homologous recombination.HEp-2 cells inoculated in vitro and streptomycin-treated mice in vivo were used to identify O157∶H7(△toxB) adherence and colonization.The results from PCR and Western blotting showed that toxB gene was successfully replaced with the kanamycin gene cassette by an allelic exchange.No differences were found between the O157∶H7(△toxB) and the parent O157∶H7 in growth feature.O157∶H7(△toxB) was two-times attenuated in adherence to HEp-2 cells.For Balb/c mice,the fecal shedding of O157∶H7(△toxB) was only 3.1×105 CFU on the third day after oral infection,whereas the parent O157∶H7 was 4.03×107 CFU.The mice infected with O157∶H7(△toxB) shed the bacteria for 11 d,while with the parent O157∶H7,the mice shed for 14 d.

相似文章

导出引用
计量
文章访问数: 11
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊

友情链接Friendly link
农业信息研究所Agricultural information institute 农业智能知识服务平台Agricultural intelligent knowledge service platform 农业学术服务平台Agricultural Knowledge Service System 中国知网CNKI 维普网VIP 万方WANFANG DATA
联系我们Contact us

电话Tell：010-82109652-808

邮箱E-mail：agrijournal@caas.cn

服务Service
关于我们About us 意见建议Opinions and suggest
版权说明Copyright
单位：中国农业科学院农业信息研究所 中国农学会 © 2017中国农业科学院农业信息研究所 ICP备案号：京ICP备09089781号-46