Cloning and bioinformatic analysis of glucosetransferase gene NtGT3 in Nicotiana tabacum

doi:10.3969/j.issn.1004-1524.2017.01.04

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

• Crop Science • Previous Articles Next Articles

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

Abstract

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

CLC Number:

S572

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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Cloning and bioinformatic analysis of glucosetransferase gene NtGT3 in Nicotiana tabacum

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