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为了研究Ⅲ型泌出效应因子在丁香假单胞大豆致病变种中的作用,利用反向PCR技术,首次从丁香假单胞大豆致病变种全基因组中克隆得到两个效应因子HopAB1和HopAF1基因的同源物,分别命名为HopAB1s和HopAF1s。生物信息学分析表明,HopAB1s基因全长是1 572 bp,编码523个氨基酸;HopAF1s基因全长是855 bp,编码284个氨基酸。即基因的登录号分别为JF826562和JF826563。保守功能区预测显示HopAB1s在N末端包含一个E3泛素连接酶功能区。将这2个基因克隆到PVX二元表达载体并转化农杆菌,利用农杆菌介导的瞬时侵染技术在本生烟中表达,发现2个效应因子均能抑制由鼠凋亡因子激发的细胞程序性死亡;将烟草疫霉接种在表达效应基因的区域,发现效应因子能促进烟草疫霉侵染烟草,因此本研究得到的两个效应因子是免疫抑制因子,为进一步研究该菌的致病机理奠定基础。
In order to study the role of type Ⅲ secretion factor in clove pseudopenic soybean pathogenicity mutant, two effect factors, HopAB1 and HopAF1, were cloned from the whole genome of pathogenic strain of clove pseudophakic soybean by reverse PCR Homologues named HopAB1s and HopAF1s respectively. Bioinformatics analysis showed that the full length of HopAB1s gene was 1 572 bp, encoding 523 amino acids. The full length of HopAF1s gene was 855 bp, encoding 284 amino acids. Namely, the accession numbers of the genes are JF826562 and JF826563, respectively. Conserved functional region prediction shows that HopAB1s contains an E3 ubiquitin ligase function domain at the N-terminus. The two genes were cloned into PVX binary expression vector and transformed into Agrobacterium tumefaciens. Agrobacterium tumefaciens-mediated transient infection was used to express in Nicotiana benthamiana. Both of the two effectors inhibited the apoptosis of cells Programmed death; Phytophthora nicotianae inoculated in the expression of effector genes in the region and found that the effect of factors can promote the tobacco Phytophthora infestans, so the two effect factors obtained in this study are immunosuppressive factors, to further study the pathogenic bacteria Mechanism laid the foundation.