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根据玉米细菌性褐斑病抗病基因的定位结果预测得到1个编码受体激酶(Receptor-like kinases,RLKs)基因Psy2,采用生物信息学方法对显性基因Psy2和隐性基因psy2编码蛋白的结构、功能和理化性质进行预测和分析。结果表明,psy2的编码氨基酸与Psy2相比共有57个氨基酸差异,包括起始密码子Met提前1位、1个精氨酸的缺失和55个氨基酸替换;两者理化性质差异较小,都被归类为不稳定蛋白;都具有信号肽,属分泌性蛋白;psy2无法形成有效跨膜结构域(Transmembrane domain,TM),但都定位于质膜;Psy2基因的进化与相应的物种进化相一致。推测psy2蛋白的氨基酸残基替换、缺失以及跨膜结构域的改变影响了信号的正常传导。通过基因枪转化法将RNA干扰表达载体导入受体HiⅡA×HiⅡB的幼胚,获得阳性转基因植株,经过q RT-PCR检测显示,Psy2a-RNAi在成株期(开花期)的干扰效果比苗期显著增强。
According to the mapping results of the resistance genes of bacterial brown spot in corn, one P RL gene encoding Receptor-like kinases (RLKs) was predicted and bioinformatics methods were used to detect the expression of the dominant gene Psy2 and the recessive gene psy2 Structure, function and physicochemical properties of prediction and analysis. The results showed that there were 57 amino acid differences between psy2 and Psy2, including 1 position of start codon, 1 arginine deletion and 55 amino acid substitutions. The difference of physicochemical properties of psy2 was Are classified as unstable proteins; all have signal peptides, which are secreted proteins; psy2 can not form a transmembrane domain (TM), but both are located on the plasma membrane; the evolution of Psy2 is consistent with the evolution of the corresponding species . It is speculated that psy2 protein amino acid residue substitutions, deletions and changes in the transmembrane domain affect the normal signal transduction. RNA interference expression vectors were introduced into immature embryos of HiⅡA × HiⅡB by gene gun transformation, and positive transgenic plants were obtained. The results of q RT-PCR showed that the interference effect of Psy2a-RNAi at the adult stage (flowering stage) Significantly enhanced.