西兰花CYP83A1基因的克隆及序列分析

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

摘要/Abstract

摘要： 为深入研究西兰花4-甲基亚磺酰丁基芥子油苷的合成代谢途径，对关键合成酶基因CYP83A1进行了克隆与生物信息学分析。根据前期西兰花转录组测序工作中获得的序列数据，同时参照GenBank数据库中拟南芥、小白菜和油菜等7种植物的CYP83A1基因CDS序列与西兰花进行比对，确定西兰花CYP83A1基因（BoCYP83A1）的CDS序列。采用RT-PCR技术对其进行了克隆，获得BoCYP83A1基因的CDS区序列，其全长为1 509 bp，编码502个氨基酸。预测该蛋白质的分子量为57.47 kD，理论等电点为 7.1，包含2个跨膜结构域，且整个序列中不含信号肽，具有一个典型的P450结构域。氨基酸同源性分析表明，西兰花与油菜、大白菜的CYP83A1的相似性较高，均为98%。首次获得BoCYP83A1的CDS序列，其登录号为KM111290。

关键词: 西兰花, CYP83A1基因, 芥子油苷, 序列分析, 抗癌活性

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

曹阳理惠, 李勇, 孔稳稳, 李晶. 西兰花CYP83A1基因的克隆及序列分析[J]. 生物技术通报, 2015, 31(2): 103-110.

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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