小麦病程相关蛋白1基因的亚细胞定位及信号肽鉴定

发布时间：2019-05-23 03:51

【摘要】：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。
[Abstract]:A wheat course-related protein 1 gene TaLr35PR1, induced by leaf rust was cloned to clarify its gene structure and expression characteristics. In this study, the signal peptide and subcellular localization of TaLr35PR1 were predicted and confirmed by experiments. The nucleic acid sequence encoded by signal peptide was cloned into yeast signal peptide trapping vector pSUC2T7M13ORI and transformed into yeast sucrase deficient strain YTK12. It was confirmed that the signal peptide of TaLr35PR1 gene had secretory function. At the same time, the subcellular localization recombinant pCamA-TaLr35PR1-GFP, was successfully constructed by enzyme digestion and ligature, and the onion epidermis cells were transiently transformed by gene gun bombardment technique. The results showed that the expression of pCamA-TaLr35PR1-GFP fusion protein was mainly extracellular, which was consistent with the prediction of subcellular localization. These results enrich the research content of wheat disease course related protein 1 gene, and provide a research basis for further exploring the biological function and mechanism of this kind of gene.
【作者单位】：河北农业大学植物保护学院/河北省农作物病虫害生物防治工程技术研究中心;
【基金】：国家自然科学基金(31501623) 河北省高等学校科学技术研究项目(QN2015171)
【分类号】：S435.12

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