小麦矮腥黑粉菌效应蛋白g11335的生物信息学分析、亚细胞定位及毒性验证

doi:10.13560/j.cnki.biotech.bull.1985.2022-0414

摘要/Abstract

摘要：

本研究重点分析小麦矮腥黑粉菌的g11335基因编码效应蛋白的生物学功能。效应蛋白g11335为GSDH家族的效应蛋白,该家族成员有影响植物体的可能性。运用多种生物信息学数据库,解析效应蛋白g11335的理化性质、保守结构域、跨膜区、亲疏水性以及信号肽,并且构建了g11335-pBin表达载体进行亚细胞定位分析。结果表明,g11335基因全长1 389 bp,共编码462个氨基酸。该效应蛋白的亚细胞定位结果表明,该蛋白质定位在细胞膜上。烟草毒性验证表明该效应蛋白无毒性。

关键词: 小麦矮腥黑粉菌, g11335, 生物信息学分析, 亚细胞定位

Abstract:

This research focused on analyzing the biological function of effector protein encoded by g11335 gene of Tilletia contraversa Kühn. Effector g11335 was an effector protein of GSDH family which may affect the plant healthy. The physicochemical properties, conserved domain, transmembrane domain, hydrophilicity and hydrophobicity of effector g11335 were analyzed using a variety of bioinformatics databases, and the expression vector g11335-pBin was constructed for subcellular localization analysis. The results showed that g11335 gene was 1 389 bp in length and encoded 462 amino acids. Subcellular localization of the effector protein showed that the protein was localized on the cell membrane. Tobacco toxicity test revealed that the effector protein was not toxic.

Key words: Tilletia contraversa Kühn, g11335, bioinformatics analysis, subcellular localization

郭志浩, 金泽鑫, 刘琦, 高利. 小麦矮腥黑粉菌效应蛋白g11335的生物信息学分析、亚细胞定位及毒性验证[J]. 生物技术通报, 2022, 38(8): 110-117.

GUO Zhi-hao, JIN Ze-xin, LIU Qi, GAO Li. Bioinformatics Analysis, Subcellular Localization and Toxicity Verification of Effector g11335 in Tilletia contraversa Kühn[J]. Biotechnology Bulletin, 2022, 38(8): 110-117.

图/表 10

表1 引物列表

Table 1 List of primers

引物名称 Primer name	引物序列 Primer sequence（5'-3'）
g11335-F	TCAGCACCAGCTAGCATCGATATGGCGCGCCAAC- TCGGCAAC
g11335-R	AACCGTTCATCGGCGGTCGACTCAAGCGGTTCCA- TCGATCGACA
g11335-YF	ATTTACGAACGATAGGGTACCATGGCGCGCCAAC- TCGGCAAC
g11335-YR	GCCCTTGCTCACCATGGATCCTCAAGCGGTTCCA- TCGATCGACA
pBin-F	ACAATCCCACTATCCTTCGC
pBin-R	AAGTCGTGCTGCTTCATGT
PGR107-F	ATAGCAGTCATTAGCACTTCCT
PGR107-R	CACTGGGGCATACTTCATGC

图1 基因g11335的PCR扩增产物

Fig. 1 Products of gene g11335 by PCR amplification M：DL2000 DNA maker

图2 效应蛋白g11335的保守结构域分析

Fig. 2 Conserved domain analysis of effector g11335

图3 效应蛋白g11335的亲疏水性图

Fig. 3 Hydrophilicity of effector g11335

图4 效应蛋白g11335信号肽预测

Fig. 4 Signal peptide prediction of effector g11335

图5 效应蛋白g11335的跨膜区预测

Fig. 5 Prediction of transmembrane region of effector g11335

图6 效应蛋白g11335的二级结构预测

Fig. 6 Secondary structure of effector g11335

图7 效应蛋白g11335的三级结构预测

Fig. 7 Tertiary structure prediction of effector g11335

图8 效应蛋白g11335亚细胞定位图 A：GFP通道下的空载体;B：在明场视野下的空载体;C：复合场下的空载体;D：GFP通道下的g11335;E：在明场视野下的g11335;F：复合场下的g11335（标尺为20 μm）

Fig. 8 Subcellular localization of effector g11335 A: Empty carrier in the GFP channels. B: Empty carrier in the bright field of vision. C: Empty carrier in the compound field. D: g11335 in the GFP channels. E: g11335 in the bright field of vision. F: g11335 in the compound field（Scale: 20 μm）

图9 效应蛋白g11335的毒性验证

Fig. 9 Toxicity validation of effector g11335

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