Cloning and Sequence Analysis of CYP83A1 Gene from Brassica oleracea var. italica

doi:10.13560/j.cnki.biotech.bull.1985.2015.02.015

Abstract

Abstract: To further research the related genes involved in 4-methylsulfinybutyl-glucosinolate biosynthesis in Brassica oleracea var. italica, the key synthetase CYP83A1gene was cloned and analyzed by bioinformatics. The sequence of CYP83A1 from the previous data of broccoli transcriptome sequencing and the sequences of CYP83A1 in seven other plants （Arabidopsis thaliana, Brassica napus and the Brassica rapa subsp. Pekinesis et.al）which obtained from GenBank were alignmented with each other using NCBI blaster, and we confirmed that the homologically conserved region of CYP83A1 gene in Brassica oleracea var. italica （BoCYP83A1）. Using RT-PCR method, the CDS region of BoCYP83A1 was obtained. Bioinformatic analysis indicated that the CDS region of BoCYP83A1 was 1 509 bp long which encoded a predicated a protein contains 502 amino acids. Protein analysis showed that BoCYP83A1 molecular weight was 57.47 kD, theoretical pI was 7.1 and contained two span membrane structures, and the BoCYP83A1 does not have signal peptide sequences, while possessed a P450 functional domains. Homology analysis of the deduced amino acids indicated that BoCYP83A1 had 98% similarity with Brassica napus and the Brassica rapa subsp. Pekinesis. The gene registration number of Brassica oleracea var. italica CYP83A1 gene is KM111290.

Key words: Brassica oleracea var. italica, CYP83A1 gene, glucosinolates, sequence analysis, antitumor activity

Cao Yanglihui, Li Yong, Kong Wenwen, Li Jing. Cloning and Sequence Analysis of CYP83A1 Gene from Brassica oleracea var. italica[J]. Biotechnology Bulletin, 2015, 31(2): 103-110.

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