绿色杜氏藻DHDDS基因生物信息学分析及表达研究

doi:10.13560/j.cnki.biotech.bull.1985.2015.07.015

摘要/Abstract

摘要： 绿色杜氏藻是一种能产生重要次生代谢产物类胡萝卜素的单细胞绿藻，脱氢多萜醇焦磷酸合成酶(dehydrodolichyl diphosphate synthase，DHDDS)是其在类胡萝卜素合成途径中的相关酶。旨在研究DHDDS基因表达与类胡萝卜素含量之间的关系。提取绿色杜氏藻总RNA，通过转录组测序获得DHDDS基因全长，对该基因进行生物信息学分析；并用不同浓度甲基茉莉酸(MeJA)处理绿色杜氏藻，采用实时定量PCR研究该基因转录差异。结果显示，绿色杜氏藻DHDDS基因全长2 211 bp，含有一个1 740 bp长的开放阅读框(ORF)，编码579个氨基酸序列。DHDDS蛋白质理论等电点为7.63，相对分子质量为62 472.7 Da。预测结果表明，DHDDS蛋白不含信号肽，也不存在跨膜区域，该蛋白定位于细胞质基质。氨基酸序列比对结果显示，绿色杜氏藻DHDDS蛋白与小球藻的同源性最高(59%)。实时定量PCR结果表明，经100 μmol/L MeJA处理的绿色杜氏藻DHDDS表达水平最高，具有极显著差异；在该浓度下，类胡萝卜素含量均高于其他浓度组。且DHDDS基因的表达与类胡萝卜素含量呈一定的相关性。绿色杜氏藻DHDDS是一种定位于细胞质基质中的酶，其与绿色杜氏藻类胡萝卜素合成途径有关。在一定浓度范围的MeJA诱导下，低浓度的MeJA对其表达起促进作用，高浓度MeJA对其表达有抑制作用，且其表达与类胡萝卜素含量呈一定的相关性。

关键词: 绿色杜氏藻, 脱氢多萜醇焦磷酸合成酶, 类胡萝卜素, 甲基茉莉酸

Abstract: Dunaliella viridis is a unicellular green alga producing valuable secondary metabolite carotenoids, and dehydrodolichyl diphosphate synthase(DHDDS)is an enzyme in the synthesis pathway of carotenoids. It was to investigate the correlation between expression of DHDDS gene in D. viridis treated with different concentrations of MeJA and the content of carotenoids. Total RNA was extracted from D. viridis and the full length of DHDDS was acquired by de-novo transcriptome sequencing, then the sequence was analyzed by bioinformatics website and software to predict the protein structure. Transcriptional levels of D. viridis treated with different concentrations of MeJA were examined by Real-time quantitative PCR(RT-qPCR). The total length of gene DHDDS was 2 211 bp, containing a 1 740 bp open reading frame(ORF)that encoded 579 amino acids. Theoretical isoelectric point of DHDDS protein was 7.63. Relative molecular mass was 62 472.7 Da. Bioinformatics prediction revealed that there was no signal peptide and transmembrane domain in DHDDS protein, and it was located in cytoplasmic matrix. Sequence alignment showed that DHDDS of D. viridis shared the highest homology of 59% with that of Chlorella variabilis. The RT-qPCR showed that the expression of DHDDS gene in D. viridis treated by 100 mol/L of MeJA was the highest with significant difference：at this concentration, the content of carotenoids was higher than other groups. There was certain correlation between the expression of DHDDS gene and the content of carotenoids. Conclusively, the DHDDS of D. viridis is an enzyme located in cytoplasmic matrix, relating to synthesis pathway of carotenoids. Within certain concentration, MeJA in lower concentration shows promoting effect on the expression, and inhibiting in higher concentration, there is a correlation between the expression and the content of carotenoids.

Key words: Dunaliella viridis, DHDDS, carotenoids contents, MeJA

潘益芳, 龚一富, 俞凯, 朱帅旗, 王何瑜. 绿色杜氏藻DHDDS基因生物信息学分析及表达研究[J]. 生物技术通报, 2015, 31(7): 101-108.

Pan Yifang, Gong Yifu, Yu Kai, Zhu Shuaiqi, Wang Heyu. Bioinformatics Analysis and Expression Study of DHDDS of Dunaliella viridis[J]. Biotechnology Bulletin, 2015, 31(7): 101-108.

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