摘要
利用Getorf、SignalP 3.0、BLASTP等生物信息学软件和PERL语言从葡萄霜霉菌基因组中预测到一条编码RXLR-EER结构域的效应蛋白候选基因,将该基因编码的蛋白命名为RXLR5。将RXLR5信号肽SP5编码序列连接到pSUC2T7M13ORI载体并转化到酵母蔗糖酶缺陷菌株YTK12后,重组菌株能成功分泌蔗糖酶,促使棉籽糖分解成单糖,因此,能在YPRAA培养基上正常生长;同时生成的单糖能与TTC反应产生红色不溶于水的氯化三苯基四氮唑。由此推测,SP5具有信号肽活性,可能在RXLR5从葡萄霜霉菌细胞内泌到胞外的过程中起重要作用。
By using bioinformatics approaches, we identified an RXLR-EER effector from Plasmopara viticola genome, RXLRS. We used the predicted signal peptide SP5 in-frame with the mature sequence of yeast invertase SUC2 in the yeast signal sequence trap vector pSUC2T7M13ORI, and the resultant construct was transferred into a yeast SUC2-minus strain. The fusion of SP5 to the SUC2 mature sequence enabled the successful secretion of invertase, thus enabling the yeast cells to hydrolyze raffinose and grow on YPRAA medium. The secretion of invertase and the hydrolysis of raffinose into monosaccharides were also confirmed with 2,3,5 - triphenyltetrazolium chloride （TFC）, which reacts with monosac- charides to form the insoluble triphenylformazan （red color）. These results suggest that SP5 might have a signal peptide activity, and play an important role in the secretion of RXLR5 out of the cell.
出处
《植物保护》
CAS
CSCD
北大核心
2016年第1期40-44,共5页
Plant Protection
基金
公益性行业(农业)科研专项(201203035)
国家现代农业产业技术体系专项资金(nycytx-30)
关键词
葡萄霜霉病菌
信号肽
酵母分泌系统
蔗糖酶活性
Plasmopara viticola
signal peptide
yeast secretion system
invertase activity
