Cloning, Expression and Vector Construction of BrSOT16 Related to Glucosinolate Synthesis in Brassica rapa ssp. chinensis L.
1 Collaborative Innovation Center of Improving Quality and Increasing Profits for Protected Vegetables in Shanxi, College of Horticulture, Shanxi Agricultural University, Jinzhong, 030801, China
2 Zhejiang Provincial Key Laboratory of Agricultural Product Quality Improvement Technology, Zhejiang Agriculture and Forestry University Biological Seed Research Center, College of Agriculture and Food Science, Zhejiang Agriculture and Forestry University, Hangzhou, 311300, China
Molecular Plant Breeding, 2020, Vol. 11, No. 29 doi: 10.5376/mpb.2020.11.0029
Received: 21 Sep., 2020 Accepted: 15 Dec., 2020 Published: 31 Dec., 2020
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This article was first published in Molecular Plant Breeding in Chinese, and here was authorized to translate and publish the paper in English under the terms of Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article:
Guo E.B., Zhu Z.J., Shi Q.Y., Ma Y.H., Yu Y.J., and Li M.L., 2020, Cloning, expression and vector construction of BrSOT16 related to glucosinolate synthesis in Brassica rapa ssp. chinensis L.,Molecular Plant Breeding, 11(29): 1-10 (doi: 10.5376/mpb.2020.11.0029)
Sulfotransferase is an enzyme that catalyzes the transfer of sulfate groups from 3’-phosphoadenosine 5’-phosphosulfate to various receptor molecules, and plays an important role in plant growth and development as well as resistance to diseases and insect pests, etc. In this study, the 1 020 bp gene sequence of BrSOT16 was obtained by homologous cloning from Brassica rapa. Bioinformatics analysis showed that BrSOT16 encoded a stable hydrophilic protein containing 339 amino acids, whose molecular mass and theoretical isoelectric point were 39.25 kD and 5.50, respectively. It was a member of the plant sulfotransferase family containing four conserved regions and having no signal peptide and transmembrane domain. Evolutionary analysis showed that BrSOT16 had the closest relationship with BcSOT16 and BnSOT16. QRT-PCR analysis showed that BrSOT16 showed an expression pattern that increased first and then decreased as the plants continued to develop, and it had the highest expression level in the leaves of pakchoi at the twelve-leaf stage. The overexpression vector of BrSOT16 was constructed by the method of homologous substitution for the biological function identification, which could provide experimental materials and technical assistance for further study on the genetic engineering of pakchoi.
Brassica rapa; Sulfotransferase; Glucosinolates; Gene clone; Gene expression; Vector construction