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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。
Using three lines of cauliflower as test material, the cauliflower BoPGIP2 gene was cloned by homologous cloning, and its expression level was analyzed. The results showed that the gene had a high similarity with the PGIP2 gene sequence in rape; ExPASy analysis showed that BoPGIP2 had a theoretical molecular weight of 37.02 kDa, an isoelectric point of 8.51 and a molecular formula of C 1672 H 26060 N 436O487 S_ 13). The coefficient of instability was 34.95, which was a stable protein with an aliphatic index of 92.48 and a GRAVY value of -0.096, indicating no significant difference in the hydrophilicity and hydrophobicity of the primary structure. Subcellular localization prediction analysis showed that this protein belonged to the signal peptide of secretory pathway. QRTPCR analysis showed that the relative expression level of BoPGIP2 gene was high in stem and leaf of the high-resistance lines of Cauliflower, In the susceptible strains, the highest relative expression level in roots and the relatively low expression level in leaves and stems were consistent with the strength of antimicrobial scleroses, suggesting that this gene may play an important role in the regulation of cauliflower’s antibacterial nuclear disease An important role.