肠炎沙门菌毒力基因在感染济宁百日鸡中的表达规律研究
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摘要
选择2日龄肠炎沙门菌阴性济宁百日鸡作为试验动物,口服灌注肠炎沙门菌。感染后第1、7、14、21、28和35天分别采集试验组、对照组盲肠内容物并进行稀释涂布,测定各样品含菌量。提取试验组个体肠道内容物中肠炎沙门菌总RNA,利用荧光定量PCR测定肠炎沙门菌毒力因子invH、sefA、lpfC在感染宿主后不同时间点盲肠内容物中的表达,探究在感染宿主中毒力基因的表达规律。结果显示:肠炎沙门菌invH、lpfC与sefA三种毒力基因感染宿主后第7、14、21、28、35天的表达量均呈下降趋势,毒力基因在感染宿主后的表达量下降,比感染前下降了3个数量级。表明感染宿主后,肠炎沙门菌毒力基因的表达受到抑制,这种抑制作用随着时间的延长而增强。
Two-day-age Salmonella enteritidis free Jiningbairi chicken was selected and inoculated with Salmonella enteritidis. The cecal content were collected on the 1 st, 7 th, 14 th, 21 st, 28 th and 35 th day post inoculation, respectively,for RNA isolation and bacteria counting. Three pathogenicity genes ofinvH, sefA and lpfC were selected to detect mRNA expression at each time point post inoculation. The results showed that the expression of three detected pathogenicity genes significantly were decreased after inoculation. The results indicated that the three pathogenicity genes were inhibited after inoculation, The repression of which were strengthened with time post inoculation.
Two-day-age Salmonella enteritidis free Jiningbairi chicken was selected and inoculated with Salmonella enteritidis. The cecal content were collected on the 1 st, 7 th, 14 th, 21 st, 28 th and 35 th day post inoculation, respectively,for RNA isolation and bacteria counting. Three pathogenicity genes ofinvH, sefA and lpfC were selected to detect mRNA expression at each time point post inoculation. The results showed that the expression of three detected pathogenicity genes significantly were decreased after inoculation. The results indicated that the three pathogenicity genes were inhibited after inoculation, The repression of which were strengthened with time post inoculation.
引文
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[3] SMATI M, MAGISTRO G, ADIBA S, et al. Strain-specific impact of the high-pathogenicity island on virulence in extra-intestinal pathogenic Escherichia coli[J]. International Journal of Medical Microbiology Ijmm, 2017, 307(1):44.
[4] ESPINOZA R A, SILVA-VALENZUELA C A, AMAYA F A, et al. Differential roles for pathogenicity islands SPI-13 and SPI-8 in the interaction of Salmonella Enteritidis and Salmonella Typhi with murine and human macrophages[J]. Biological Research, 2017, 50(1):5.
[5] MORITA A, TAI A, ITO H,et al. Proanthocyanidins in an astringent persimmon inhibit Salmonella pathogenicity Island 1(SPI1)secretion[J]. Journal of the Science of Food&Agriculture, 2016, 96(5):1798-1802.
[6]余剑岚.鼠伤寒沙门氏菌适应宿主环境的蛋白质组学研究[D].武汉:武汉大学,2011.
[7]王俊红,刘高生,牛钟相,等.沙门氏菌invH基因的序列分析与分子检测[J].江西农业大学学报,2008(30):324-327.
[8]王晓春,焦扬,程瞾,等.肠炎沙门菌lpfC基因缺失株的构建及基本生物学特性研究[J].生物技术,2015,25(3):256-274.
[9]董立伟.肠炎沙门氏菌鞭毛素fliC基因缺失突变株的构建及相关功能性分析[D].扬州:扬州大学,2014.
[10]王小龙.沙门氏菌invH基因亚单位疫苗和C3d分子佐剂核酸疫苗的制备与免疫效果研究[D].泰安:山东农业大学,2011.
[11]陈静.利用SCOTS技术筛选肠炎沙门菌在不同源巨噬细胞内的转录序列[D].扬州:扬州大学,2013.
[12]朱春红,孟霞,姚丰华,等.肠炎沙门氏菌SEF14菌毛亚单位sefA和sefD基因缺失株的致病性研究[J].中国预防兽医学报,2012,34(10):786-789.
[13] EDWARD R A, SCHIFFERLI D M, MALOY S R. A role for Salmonella fimbriae in intraperitoneal infections[J].Proceedings of the National Academy of Sciences of the United States of America, 2000, 97(3):1258-1262.
[14] CLOUTHLER S C, COLLINSON S K, KAY W W. Unique fimbriae-like structures encoded by sefD of the SEF14fimbrial gene cluster of Salmonella Enteritidis[J]. Molecular Microbiology, 2010, 12(6):893-903.