Cloning and bioinformatics analysis of VvGST1 from Munage tab.grapes

doi:10.6048/j.issn.1001-4330.2023.08.013

Xinjiang Agricultural Sciences ›› 2023, Vol. 60 ›› Issue (8): 1922-1930.DOI: 10.6048/j.issn.1001-4330.2023.08.013

• Horticultural Special Local Products · Agricultural Product Processing Engineering • Previous Articles Next Articles

Cloning and bioinformatics analysis of VvGST1 from Munage tab.grapes

WANG Man¹(), ZHANG Zheng², Yilidana Dilixiati¹, WU Bin²()

1. College of Food and Pharmaceutical Sciences, Xinjiang Agricultural University, Urumqi 830052, China
2. Institute of Agro-products Storage and Processing, Xinjiang Academy of Agricultural Sciences/Xinjiang Key Laboratory of Processing and Preservation of Agro- products, Urumqi 830091,China

Received:2022-12-01 Online:2023-08-20 Published:2023-08-14
Correspondence author: WU Bin(1973-), male, native place: Tacheng, Xinjiang.Professor, research field: Storage and processing of agricultural products, (E-mail)42042615@qq.com.
Supported by:
National Natural Science Foundation of China(U2003213);National Natural Science Foundation of China(31860460);Xinjiang Uygur Autonomous Region Regional Collaborative Innovation Project(2021E01005)

木纳格葡萄谷胱甘肽-S-转移酶VvGST1基因克隆与序列分析

王曼¹(), 张政², 伊丽达娜·迪力夏提¹, 吴斌²()

1.新疆农业大学食品科学与药学学院,乌鲁木齐 830052
2.新疆农业科学院农产品贮藏加工研究所/新疆农产品加工与保鲜重点实验室,乌鲁木齐 830091

通讯作者: 吴斌(1973-),男,新疆塔城人,研究员,博士(后),硕士生/博士生导师,研究方向为农产品贮藏与加工,(E-mail)42042615@qq.com
作者简介:王曼(1995-),女,新疆温泉人,硕士研究生,研究方向为农产品贮藏与加工,(E-mail)1803348774@qq.com
基金资助:
国家自然科学基金项目(U2003213);国家科学自然基金项目(31860460);新疆维吾尔自治区区域协同创新专项(2021E01005)

Abstract

Abstract:

【Objective】 To clone and study the sequence characteristics of the glutathione-S-transferase, (GST) gene from the Vitis vinifera L.cv ‘Munage’ table grapes in the hope of laying a foundation for the disease resistance and function of this gene in grape fruit.【Methods】 Full-length VvGST1 gene cDNA was isolated from grape fruit according to the existed ESTs sequence,and 3' and 5' end of the RACE primers were designed by using RT-PCR and RACE.【Results】 A variety of bioinformatics analysis methods were used to predict and analyze the gene, and the results showed that the ORF (Open Reading Frame) length was 719 bp, which encoded a 221 amino acid polypeptide.The VvGST1 gene was forecasted and the relative molecular mass of encoding the protein was 25.55 kDa, the theoretical isoelectric point was 6.32, the atomic composition was C₁₁₇₈H₁₇₉₉N₂₉₃O₃₂₁S₁₁.Its instability coefficient was 39.22, showing it was astable hydrophilic protein without signal peptide or transmembrane structure.It might exist in the cell matrix and was a typical matrix protein.Through analyzing the evolutionary tree, the amino acid sequence of VvGST1 was clustered with cotton, peach, prunus, cherry, plum, chestnut and other plants, among which it was closely related to cotton.【Conclusion】 The full-length coding region of VvGST1 gene in Vitis vinifera L.cv ‘Munage’ table grapes is successfully obtained,and the structural characteristics of the sequence are confirmed.

Key words: grape; cloning; glutathione-S-transferase; VvGST1 gene; sequence analysis

摘要：

【目的】克隆木纳格葡萄果实中谷胱甘肽-S-转移酶(glutathione-S-transferase,GST)基因VvGST1全长并研究其序列特征,为该基因在葡萄果实抗病作用和功能的研究奠定基础。【方法】根据已有的基因序列,设计3'和5'端RACE引物,利用RT-PCR和RACE技术克隆VvGST1基因cDNA全长。【结果】该基因序列全长719 bp,均为开放阅读框(ORF),共编码221个氨基酸。该基因编码蛋白相对分子质量为25.55 kDa;理论等电点pI值为6.32;分子式为C₁₁₇₈H₁₇₉₉N₂₉₃O₃₂₁S₁₁;不稳定指数为39.22;该蛋白质是稳定的亲水性蛋白,不含跨膜结构域,没有预测到信号肽,可能存在于细胞基质中,是典型的基质蛋白。VvGST1氨基酸序列与棉花、桃、西梅、樱桃、李子、板栗等植物聚为一类,其中与棉花属亲缘关系最近。【结论】获得了木纳格葡萄谷胱甘肽-S-转移酶VvGST1基因全长编码序列,探明了该序列的结构特征。

关键词: 葡萄, 克隆, 谷胱甘肽-S-转移酶, VvGST1基因, 序列分析

CLC Number:

S663.1

WANG Man, ZHANG Zheng, Yilidana Dilixiati, WU Bin. Cloning and bioinformatics analysis of VvGST1 from Munage tab.grapes[J]. Xinjiang Agricultural Sciences, 2023, 60(8): 1922-1930.

王曼, 张政, 伊丽达娜·迪力夏提, 吴斌. 木纳格葡萄谷胱甘肽-S-转移酶VvGST1基因克隆与序列分析[J]. 新疆农业科学, 2023, 60(8): 1922-1930.

Figures/Tables 10

Fig.1 PCR amplification of electrophoresis chart sequencing peak diagram and comparison resultsof VvGST1 gene in grape

Fig.2 Multiple sequence alignment of amino acids of the VvGST1 proteins isolated to other species

Fig.3 Phylogentic relationship of amino acid sequences between VvGST1 and other GST proteins

Fig.4 Amino acid composition of protein encoded and hydrophobicity by the VvGST1 gene

Fig.5 Hydrophilicity analysis of VvGST1 protein

Fig.6 Predictive analysis of VvGST1 signal peptide

Fig.7 Prediction of VvGST1 protein transmembrane domain

Tab.1 Prediction of VvGST1 subcellular location

亚细胞定位 Subcellular location	分值Score
亚细胞定位 Subcellular location	LocDB	PotLocDB	Neural Nets	Pentamers	Integral
细胞核 Nuclear	0.0	0.0	0.00	0.00	0.05
质膜 Plasma membrane	0.0	0.0	0.96	0.00	0.08
胞外 Extracellular	0.0	0.0	0.96	0.00	0.00
细胞质 Cytoplasmic	10.0	3.0	0.00	2.44	8.64
线粒体 Mitochondrial	0.0	0.0	0.00	2.42	0.08
内质网 Endoplasm. retic	0.0	0.0	0.00	0.80	0.14
过氧化物酶体 Peroxisomal	0.0	0.0	0.96	0.0	0.00
高尔基体 Golgi	0.0	0.0	0.13	0.26	0.00
叶绿体 Chloroplast	0.0	0.0	0.00	0.03	0.67
液泡 Vacuolar	0.0	0.0	0.00	0.00	0.34

Fig.8 Prediction of the secondary structure of VvGST1 protein

Fig.9 The tertiary structure prediction of VvGST1 protein

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Cloning and bioinformatics analysis of VvGST1 from Munage tab.grapes

木纳格葡萄谷胱甘肽-S-转移酶VvGST1基因克隆与序列分析

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