绿色杜氏藻GGR基因生物信息学分析及表达研究  

Induced expression and bioinformatic analysis of GGR gene in Dunaliella viridis

作　　者：张燕[1] 龚一富[1] 景丹丹[1] 朱帅旗 俞凯[1] 

机构地区：[1]宁波大学海洋学院,浙江宁波315211

出　　处：《宁波大学学报（理工版）》2017年第1期35-41,共7页Journal of Ningbo University：Natural Science and Engineering Edition

基　　金：浙江省科技厅重点科技创新团队项目(2012R10029-07;2010R50029);宁波市科技攻关项目(2013C10018);浙江省大学生科技创新活动计划暨新苗人才计划项目(2014R405050);宁波大学优秀学位论文培育基金(py2014020)

摘　　要：采用Illumina HiSeqTM 2000高通量测序方法获得绿色杜氏藻GGR基因(Gen Bank序号:KX100480)cDNA全长序列,研究了甲基茉莉酸(Me JA)对绿色杜氏藻GGR基因表达的影响.测序结果表明,绿色杜氏藻GGR基因cDNA全长2 356 bp,开放阅读框1 539 bp,编码512个氨基酸,分子量为52.296 1 k Da,理论等电点为6.54.序列分析结果表明,该蛋白具有保守的Chl P结构域,为可溶性蛋白,具有跨膜区域,无信号肽.Target P 1.1 Server预测结果表明,该蛋白可能定位于线粒体.系统进化树结果表明,绿色杜氏藻GGR蛋白较其他植物的GGR蛋白亲缘关系较远.RT-PCR结果表明,100μmol·L-1 Me JA可显著地提升绿色杜氏藻GGR基因的表达(P<0.01).同时,绿色杜氏藻叶绿素和类胡萝卜素含量达到最高,说明GGR基因与绿色杜氏藻叶绿素和类胡萝卜素的合成存在一定关系.Geranylgeranyl reductase （GGR） is an important enzyme in the upstream of plant tocopherols and chlorophyll biosynthetics pathway. In this study, the full length cDNA of GGR gene （GenBank accession number KX100480） in Dunaliella viridis is obtained by HiSeqTM Illumina 2000. Transcriptional levels of GGR gene with the treatment of Methyl Jasmonate （MeJA） are examined by Real-Time quantitative PCR （RT-PCR）. The results show that the total length of GGR gene is 2 356 bp, containing an open reading frame of 1 539 bp encoding 512 amino acids, relative molecular weight is 52.296 1 kD and isoelectric point is 6.54. Bioinformatics prediction reveals that the protein contains a transmembrane domain without signal peptide. TargetP 1.1 Server prediction results suggest that it is located in mitochondria. Phylogenetic tree results indicate that the protein sequence of GGR in D. viridis is related loosely to other species. RT-PCR result demonstrates that the expression of GGR gene in D. viridis is promoted under the treatment of low concentration of MeJA and limited in high concentration of MeJA. When treated with 100 μmol.L-1 MeJA, the expression of GGR gene manifests the significant difference in the control group （P〈0.01）. And the content of chlorophyll and carotenoids show the same trend. The result implies that the GGR gene is to certain extent correlated with the synthetic of chlorophyll and carotenoids.

关 键 词：绿色杜氏藻 GGR 甲基茉莉酸 叶绿素 类胡萝卜素 

分 类 号：Q786[生物学—分子生物学]