向日葵锈菌弹性金属蛋白酶基因克隆及生物信息学分析

doi:10.7668/hbnxb.201751261

摘要/Abstract

摘要： 为了研究向日葵锈病的致病机制，将向日葵锈菌转录组中表达量较高的一个基因（KU994904）中成熟部分进行生物信息学分析及基因克隆。利用Interpro对蛋白ORF区域进行分析，确定其成熟区域，通过生物信息学相关软件对其进行结构预测及预测分析，从向日葵锈菌中提取总RNA，并利用RT-PCR方法扩增此蛋白酶成熟基因并克隆。结果显示：该蛋白酶成熟基因序列全长1 335 bp，编码445个氨基酸，分子质量49.5 ku，其与M36家族中5条序列具有较高相似性，并发现同胞外金属蛋白酶基因序列相似度达到53.77%。因此，推断其为弹性金属蛋白酶。

关键词: 向日葵锈菌, 弹性金属蛋白酶, 克隆, 生物信息学分析

Abstract: In order to study the pathogenic mechanism of Puccinia helianthi Schw, bioinformatics method was used analyze the mature part of KU994904 gene that had a high expression level in the transcriptome of Puccinia helianthi Schw. The protein ORF region was analyzed by Interpro which can determine its mature region. Using the bioinformatical databases and softwares, its function was analyzed and predicted. Total RNA was extracted from Puccinia helianthi Schw and used to amplify the mature gene of the metalloproteinase by RT-PCR. Finally, the gene sequence of mature protein was cloned. The results showed that the sequence full-length of the gene was 1 335 bp, coding 445 amino acids. And the protein molecular weight was 49.5 ku, it was found that the gene had high similarity with five sequences of M36 family and the gene had 53.77% similarity with the extracellular metalloproteinase (mep). So it was deduced that the gene coded an elastinolytic metalloproteinase.

Key words: Puccinia helianthi Schw, Elastinolytic metalloproteinase, Cloning, Bioinformatics analysis

中图分类号:

Q78
S432.01

李鑫淳, 宋阳, 路妍, 景岚. 向日葵锈菌弹性金属蛋白酶基因克隆及生物信息学分析[J]. 华北农学报, 2019, 34(3): 186-193. doi: 10.7668/hbnxb.201751261.

LI Xinchun, SONG Yang, LU Yan, JING Lan. Cloning and Bioinformatics Analysis of Elastinolytic Metalloproteinase Gene from Puccinia helianthi Schw[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2019, 34(3): 186-193. doi: 10.7668/hbnxb.201751261.

http://www.hbnxb.net/CN/Y2019/V34/I3/186

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