华北农学报2012，Vol.27Issue(1)：57-62,6.
应用抑制性差减杂交筛选副猪嗜血杆菌4型菌株可能毒力基因
Putative Virulence Associated Genes of Haemophilus parasuis Type 4 Strains Identified by Suppression Subtractive Hybridization
摘要
Abstract
The objective of this paper is to find putative virulent genes of Haemophilus parasuis ( HPS). Suppression subtractive hybridization ( SSH) was used to construct the subtracted library between virulent strain SW124 (HPS serovar 4 reference strain) and non-virulent strain H465 ( HPS serovar 11 reference strain). Using Southern hybridization and PCR to screen the differentially expressed genes. Seven clones were identified containing fragments that were present in the SW124 genome while absent in the H465 genome and the DNA sequences of tttese fragments were determined. These DNA fragments involved putative virulence associated genes encoded 7 proteins, including transport systems periplasmic components protein,phage recombination protein, phosphoribosylformylglyci-namidine synthase, ribosomal-protein-alanine N-acetyltransferase, ABC-type nitrate/sulfonate/bicarbonate trans-portsystems periplasmic components protein, hypothetical protein and unknown. Based on PCR analysis of the 9 HPS serovar strains, these DNA fragments were found in most virulent serovars(4,5 ,12 ,14 ,15 ) ,but absent in the non-virulent serovars(3 ,11 ). The subtracted library of HPS SW124 and H465 was successfully constructed using SSH technique and seven specific genomic fragments were obtained. In additon,most of these fragments could be detected in other serotypes HPS virulent strains with different distribution.关键词
副猪嗜血杆菌/抑制性差减杂交/毒力基因Key words
Haemophilus parasuis/Suppression subtractive hybridization( SSH) / Virulence associated gene分类
生物科学引用本文复制引用
李淼,李春玲,宋帅,杨冬霞..应用抑制性差减杂交筛选副猪嗜血杆菌4型菌株可能毒力基因[J].华北农学报,2012,27(1):57-62,6.基金项目
广东省农业科技重大专项(2009A020101006) （2009A020101006）
广州市农业科技重大专项(2009A1-E041) （2009A1-E041）
广东省农科院兽医研究所所长基金 （）
