烟草糖基转移酶基因NtGT3的克隆与生物信息学分析

doi:10.3969/j.issn.1004-1524.2017.01.04

浙江农业学报 ›› 2017, Vol. 29 ›› Issue (1): 23-30.DOI: 10.3969/j.issn.1004-1524.2017.01.04

烟草糖基转移酶基因NtGT3的克隆与生物信息学分析

任学良^{1, *}, 李立芹², 许力², 郭玉双¹, 鲁黎明^{2, *}

1.贵州省烟草科学研究院烟草行业分子遗传重点实验室,贵州贵阳 550081;
2.四川农业大学农学院,四川成都 611130

收稿日期:2016-06-09 出版日期:2017-01-15 发布日期:2017-02-23
通讯作者: 任学良,E-mail:renxuel@126.com;鲁黎明,E-mail:renxuel@126.com
作者简介:任学良(1976—),男,山西孝义人,研究员,博士,主要从事烟草育种及生物技术研究。E-mail:renxuel@126.com
基金资助:
国家烟草专卖局科技项目(110201401007(JY-07)); 贵州省烟草科学研究院科技项目(烟草基因遗传转化及表型分析)

Cloning and bioinformatic analysis of glucosetransferase gene NtGT3 in Nicotiana tabacum

REN Xueliang¹, LI Liqin², XU Li², GUO Yushuang¹, LU Liming^{2, *}

1. Key Laboratory of Molecular Genetics of CNTC, Guizhou Academy of Tobacco Sciences, Guiyang 550081, China;
2. College of Agronomy, Sichuan Agricultureal University, Chengdu 611130, China

Received:2016-06-09 Online:2017-01-15 Published:2017-02-23

摘要/Abstract

摘要： 为了给烟草糖基转移酶的生物学功能研究奠定基础,根据在GenBank上登录的烟草糖基转移酶基因NtGT3的CDS序列,以烟草栽培品种K326为材料,采用PCR的方法,对目的基因进行克隆,并测序。根据测序的结果,采用各类分析软件,对所获得的DNA片段进行生物信息学分析。结果通过PCR的方法,成功克隆了烟草糖基转移酶基因NtGT3的全长CDS序列。生物信息学分析结果表明,该基因CDS全长1 449 bp,所编码的蛋白质包含482个氨基酸。其分子量为54.15 ku,等电点为5.53。在二级结构上,α-螺旋和无规则卷曲为蛋白的主要结构形式。在三级结构上,具有42%的α-螺旋以及15%的β-折叠。NtGT3蛋白是一个亲水性蛋白质,含有2个糖基化位点,不含信号肽。NtGT3蛋白广泛分布于细胞中,在细胞膜、微体、高尔基体及内质网膜的分布分别达到了79%、31.9%、30%及20%。功能结构域分析结果证实,该蛋白包含有糖基转移酶家族的功能保守域PLN03015,属于GTB类型的糖基转移酶超级家族。同源性分析结果表明,NtGT3蛋白与枸杞、睡茄、甘薯、可可、猕猴桃、杨树、葡萄、野生大豆及拟南芥等的糖基转移酶有较高的序列相似性。NtGT3基因的成功克隆,为该基因功能的深入研究提供了理论支撑。

关键词: 烟草, 糖基转移酶, 基因

Abstract: In order to lay the foundation for the investigation of the biological function of tobacco glycosyltransferase, the CDS sequence of tobacco glycosyltransferase NtGT3 (accession number BAB88934.1) was applied, and specific primers were designed accordingly. For the cloning of NtGT3, total tobacco RNA was extracted from the leaf of cultivar K326, and RT-PCR was performed. Finally, full length CDS sequence of NtGT3 was successfully obtained. Bioinformatics analysis showed that the full-length CDS of NtGT3 was 1 449 bp, encoding protein contains 482 amino acids. Its molecular weight is 54.15 ku, and the isoelectric point is 5.53. In the secondary structure, the alpha helix and random coil are the main structural forms of the protein. The tertiary structure of this protein includes 42% alpha helix and 15% beta folding. NtGT3 is a hydrophilic protein, containing two glycosylation sites and does not contain signal peptides. Protein NtGT3 is widely distributed in the cells and the distribution rate was 79%, 31.9%, 30% and 20% respectively in the cell membrane, bodies, Golgi and the endoplasmic reticulum. Functional domain analysis results confirmed that the protein contains a functional conserved domain PLN03015, which belonged to the GTB type of glycosyltransferase superfamily. The results of homology analysis showed that NtGT3 protein shares high sequence similarity with that of Lycium barbarum, Withania somnifera, Ipomoea batatas, Theobroma cacao, Actinidia deliciosa, Populus trichocarpa, Vitis vinifera, Glycine soja, and Arabidopsis thaliana. The successful cloning of NtGT3 gene provides a theoretical support for the further study of the function of this gene.

Key words: Nicotiana tabacum, glucosyltransferase, gene

中图分类号:

S572

任学良, 李立芹, 许力, 郭玉双, 鲁黎明. 烟草糖基转移酶基因NtGT3的克隆与生物信息学分析[J]. 浙江农业学报, 2017, 29(1): 23-30.

REN Xueliang, LI Liqin, XU Li, GUO Yushuang, LU Liming. Cloning and bioinformatic analysis of glucosetransferase gene NtGT3 in Nicotiana tabacum[J]. , 2017, 29(1): 23-30.

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烟草糖基转移酶基因NtGT3的克隆与生物信息学分析

Cloning and bioinformatic analysis of glucosetransferase gene NtGT3 in Nicotiana tabacum

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