花椰菜BoPGIP2基因的克隆与表达水平分析

Cloning and Expression Analysis of BoPGIP2 Gene From Brassica oleracea

以3个品系花椰菜为试材,采用同源克隆的方法,克隆了花椰菜BoPGIP2基因,并对表达水平进行分析。结果表明:该基因与油菜的PGIP2基因序列有较高的相似性;ExPASy分析发现BoPGIP2理论分子量为37.02kDa,等电点为8.51,分子式为C_(1672)H_(2604)N_(436O487)S_(13),不稳定系数为34.95,为稳定蛋白,脂肪族指数为92.48,GRAVY值为-0.096,表明其在一级结构上的亲水性、疏水性差异不显著。亚细胞定位预测分析表明,此蛋白属于分泌途径信号肽蛋白;qRTPCR分析发现BoPGIP2基因在花椰菜高抗菌核病品系、中抗菌核病品系中均是茎和叶中相对表达量较高,而在易感菌核病品系中则是根中相对表达量最高,叶和茎中相对表达量较低,这与抗菌核病强弱具有一致性,推测此基因很可能在调节花椰菜的抗菌核病中起着重要作用。

Taking 3 strains Brassica napus as materials,using homologous cloning method,the PGIP2 gene of Brassica oleracea was cloned and the expression level was analysed.The results showed that BoPGIP2 had high similarity with PGIP2 gene of Brassica napus.The ExPASy analysis showed that molecular weight of BoPGIP2 was 37.02 kDa,theoretical pI was 8.51.The instability index（II）was 34.95,and this showed the protein was stable.Grand average of hydropathicity was-0.096,so it was not a hydrophilic protein.The subcellular localization analysis indicated that BoPGIP2 was a signal peptide of secretory pathway.qRT-PCR found that BoPGIP2 has high expression in stems and leaves in both high resistant varieties and resistant varieties,whereas BoPGIP2 expressed highly in root in susceptible varieties,it was consistent with their resistance to Sclerotinia sclerotiorum.Therefore,it might play an important role in the regulation of resistance to Sclerotinia sclerotiorumof Brassica oleracea.