国家自然科学基金、湖南省自然科学基金
The National Natural Science Foundation of China, The Natural Science Foundation of Hunan Province
角鲨烯含量是高品质植物食用油评判标准的重要指标之一。鲨烯合酶是角鲨烯合成直接相关的上游调控关键酶。本研究在已构建的油茶种仁转录组数据库基础上,设计特异引物,采用RACE技术获得油茶鲨烯合酶基因的全长cDNA克隆,命名为CoSQS(GenBank登录号为JX914592)。生物信息学分析结果表明:该基因全长1554 bp,开放阅读框为1245 bp,编码415个氨基酸残基,CoSQS蛋白为弱碱性非分泌型蛋白,具有2个明显的跨膜区和2个角鲨烯和番茄红素合成酶的特异信号区。与柿DkSQS同源蛋白亲缘关系最近,属于疏水性蛋白。亚细胞定位实验结果显示该基因定位于叶绿体。实时荧光定量PCR分析表明,5月-10月间,油茶种仁中CoSQS基因表达量呈现先上升后下降的趋势,转录最高峰在9月下旬。通过关联分析明确CoSQS基因表达量与角鲨烯含量密切相关。
Squalene content is one of evaluation standards for high quality edible oil. Squalene synthase is the directly upstream regulation key enzyme for squalene synthesis. In this study, based on the transcriptome database of Camellia oleifera seed kernel, the specific primers were designed and full-length cDNA clone was obtained by RACE. The cDNA was named CoSQS (GenBank accession No. JX914592). The results of bioinformatics analysis showed that: the cDNA full-length was 1554 bp with a 1245 bp-open reading frame, the gene encoded 415 amino acid residues. CoSQS protein was alkalescent non-secreted protein with two transmembrane regions and two specific signal area of squalene and phytoene synthase. CoSQS protein had the closest relationship with Diospyros kaki DkSQS and belonged to a hydrophobic protein. Real time fluorescent quantitative PCR analysis showed that: during May to October, the CoSQS expression firstly up-regulation and later down-regulation. The transcription peak was in late September. CoSQS gene expression was closely related with squalene content ,but no significant correlation with oil yield.
曾艳玲,张党权,曾晓峰,等.油茶种仁鲨烯合酶基因的分子特性与表达分析[J].植物遗传资源学报,2016,17(4):748-755.
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