Li Qiuchun, Xu Yaohui, Huang Jinlin, Jiao Xinan
Jiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou 225009, China.
Wei Sheng Wu Xue Bao. 2010 Nov;50(11):1545-9.
Cloning of ipaJ gene from Salmonella pullorum C79-13, and identification of expressed IpaJ protein as an immunogen of the pathogen.
With suppression subtractive hybridization (SSH) between Salmonella pullorum strain C79-13 (tester) and Salmonella enteritidis strain 50041 (driver), three subtracted fragments PEA3, PE31 and PE44 showed high homology with ipaJ in plasmid pSFD10 of Salmonella choleraesuis C500. The three subtracted sequences were spliced together into the whole sequence of ipaJ in Salmonella pullorum. Then the ipaJ gene was amplified from Salmonella pullorum by polymerase chain reaction (PCR) and cloned into prokaryotic expressive vector pET-30a(+). Western-blot was used to identify it as an immunogen. The distribution of the gene was also detected in Salmonella pullorum isolates.
The ipaJ gene cloned from Salmonella pullorum was 840 bp, and the expressed fusion protein was 37 kDa. Specific reaction was found between Salmonella pullorum positive serum and expressed protein by Western-blot assay, confirming its identification as an immunogen of Salmonella pullorum. The PCR results showed that the gene exists in all Salmonella pullorum strains.
The ipaJ gene from Salmonella pullorum was first reported and cloned, and the expressed IpaJ protein was confirmed as an immunogen of Salmonella pullorum.
从鸡白痢沙门氏菌C79 - 13中克隆ipaJ基因，并鉴定表达的IpaJ蛋白作为该病原菌的免疫原。
利用鸡白痢沙门氏菌C79 - 13（测试组）与肠炎沙门氏菌50041（驱动组）之间的抑制性消减杂交（SSH），三个消减片段PEA3、PE31和PE44与猪霍乱沙门氏菌C500质粒pSFD10中的ipaJ具有高度同源性。将这三个消减序列拼接成鸡白痢沙门氏菌ipaJ的完整序列。然后通过聚合酶链反应（PCR）从鸡白痢沙门氏菌中扩增ipaJ基因，并克隆到原核表达载体pET - 30a(+)中。采用Western - blot鉴定其为免疫原。还检测了该基因在鸡白痢沙门氏菌分离株中的分布情况。
从鸡白痢沙门氏菌中克隆的ipaJ基因长度为840 bp，表达的融合蛋白为37 kDa。Western - blot分析显示鸡白痢沙门氏菌阳性血清与表达蛋白之间有特异性反应，证实其为鸡白痢沙门氏菌的免疫原。PCR结果表明该基因存在于所有鸡白痢沙门氏菌菌株中。
首次报道并克隆了鸡白痢沙门氏菌的ipaJ基因，且证实表达的IpaJ蛋白为鸡白痢沙门氏菌的免疫原。
