| 小麦矮腥黑粉菌g9890基因编码效应蛋白的生物信息学分析及亚细胞定位 |
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| 引用本文：许停,刘太国,高利,陈万权.小麦矮腥黑粉菌g9890基因编码效应蛋白的生物信息学分析及亚细胞定位.植物保护学报,2023,50(4):866-872 |
| DOI：10.13802/j.cnki.zwbhxb.2023.2022030 |
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| 作者 | 单位 | E-mail | | 许停 | 甘肃农业大学植物保护学院, 兰州 730070
中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193 | | | 刘太国 | 中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193 | | | 高利 | 中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193 | lgao@ippcaas.cn | | 陈万权 | 甘肃农业大学植物保护学院, 兰州 730070
中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193 | wqchen@ippcaas.cn |
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| 中文摘要:为明确小麦矮腥黑粉菌Tilletia controversa g9890基因编码效应蛋白的生物学功能，根据小麦矮腥黑粉病菌转录组测序结果，筛选出效应蛋白g9890，通过PCR技术获得g9890基因cDNA的全长序列，并对其进行生物信息学以及亚细胞定位分析。多种生物信息学数据库分析表明，g9890基因全长为1 038 bp （包括终止密码子），共编码345个氨基酸，相对分子质量为37 353.32，理论等电点为5.02。g9890效应蛋白不稳定系数为15.94，疏水性指数为-0.312，是一种亲水性且稳定的蛋白。将g9890基因与pbin-GFP载体重组，利用冻融法转化至根癌农杆菌Agrobacterium tumefaciensGV3101中，将其注入烟草进行瞬时表达分析，并通过共聚焦激光显微镜观测该基因的定位状况，结果显示，g9890定位在细胞膜和细胞核上。 |
| 中文关键词:小麦矮腥黑粉菌 g9890 基因克隆 生物信息学分析 亚细胞定位 |
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| Bioinformatics analysis and subcellular localization of g9890 gene product of dwarf bunt fungus Tilletia controversa |
| Author Name | Affiliation | E-mail | | Xu Ting | College of Plant Protection, Gansu Agricultural University, Lanzhou 730070, Gansu Province, China
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China | | | Liu Taiguo | State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China | | | Gao Li | State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China | lgao@ippcaas.cn | | Chen Wanquan | College of Plant Protection, Gansu Agricultural University, Lanzhou 730070, Gansu Province, China
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China | wqchen@ippcaas.cn |
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| Abstract:To clarify the biological function of the effector protein encoded by the g9890 gene of dwarf bunt fungus Tilletia controversa. Based on the sequencing data of transcriptome of the wheat dwarf smut, the effector protein gene g9890 was selected and amplified by reverse transcription polymerase chain reaction, and then bioinformatic analysis and subcellular localization experiments were performed. The results showed that the full-length of g9890 gene was 1 038 bp which encode 345 amino acids with the predicted molecular mass of 37 353.32 and the pI of 5.02. The g9890 protein had an instability coefficient of 15.94 with a hydrophobicity index of -0.312, showing that it was a hydrophilic and labile protein. Subsequently, the g9890 gene was ligated with the pBin-GFP vector, and the recombinant plasmids were transformed into Agrobacterium tumefaciens GV3101 and injected into tobacco for transient expression. Subcelular localization of the gene was observed under confocal laser microscope. The results exhibited that the g9890 protein was localized on the cell membrane and in the nucleus. |
| keywords:Tilletia controversa g9890 gene cloning bioinformatic analysis subcellular localization |
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