小麦病程相关蛋白1基因的亚细胞定位及信号肽鉴定

前期研究克隆获得了一个叶锈菌诱导的小麦病程相关蛋白1基因TaLr35PR1,明确了其基因结构和表达特征。本研究首先预测TaLr35PR1的信号肽和亚细胞定位情况,进而通过试验予以佐证。将信号肽编码的核酸序列克隆、连接到酵母信号肽诱捕载体pSUC2T7M13ORI上,并转化到酵母蔗糖酶缺陷菌株YTK12中,明确了TaLr35PR1基因的信号肽具有分泌功能。同时,利用酶切连接的方法成功构建亚细胞定位重组体pCamA-TaLr35PR1-GFP,利用基因枪轰击技术瞬时转化洋葱表皮细胞。结果表明,pCamA-TaLr35PR1-GFP融合蛋白主要在细胞外表达,与亚细胞定位预测的结果一致。这些研究结果丰富了小麦病程相关蛋白1基因的研究内容,为进一步探索该类基因的生物学功能及其作用机制提供了研究基础。

Identification of the signal peptide and subcellular localization of pathogenesis-related protein 1 gene from wheat

In previous research,a pathogenesis-related protein gene named TaLr35PR1 was ob tained from wheat induced by leaf rust,and its structure characterization and expression profiling was identified.In this study,we predicted the signal peptide and subcellular localization of TaLr35PR1,and then proved it using experiments.The predicted signal peptide was cloned and linked to the trap vector pSUC2T7M130RI,and then transformed to yeast invertase deficient strain YTK12.The results showed that the signal peptide of TaLr35PR1 have the secretion activity.Meanwhile,subcellular location recombination vector pCamA-TaLr35PR1-GFP was constructed using enzyme digestion and ligation.The onion epidermal cells via particle bombardment showed that pCamA-TaLr35PR1-GFP fusion protein mainly distributed in extracellular environment.These results enriched the research of the pathogenesis-related protein 1 gene and provide the basis for the biological function and its mechanism.