甘蓝型油菜水苏糖合成酶基因克隆及表达分析

Cloning and Expression Analysis of Stachyose Synthase Gene in Brassica napus L.

以甘蓝型油菜品种‘德油5号’为试材,采用RT-PCR和RACE方法首次从油菜中获得水苏糖合成酶基因cDNA全长序列,命名为BnSTS,GenBank登录号为HQ285880.1。该cDNA全长3 210bp,包含一个长为2 619bp的开放阅读框,编码873个氨基酸。通过构建原核表达载体,成功表达出预期蛋白。同源性比对结果显示,该基因编码蛋白与拟南芥STS相似性最高为82.08%。荧光定量PCR结果显示,基因表达水平与油菜种子发育过程中水苏糖含量变化具有相关性,与种子脱水耐性的获得具有时间一致性。该基因可能在种子脱水耐性形成机制中具有一定的作用。

In this study,a full-length cDNA sequence of stachyose synthase gene was obtained from Brassica napus cultivar ＇Deyou 5＇ for the first time using RT-PCR and RACE,named BnSTS（GenBank accession No.HQ285880.1）.The full length of BnSTS is 3 210 bp,and the ORF is 2 619 bp,encoding 873 amino acids.Prokaryotic expression showed the recombinant plasmid can express the prospective proteins.Homology analysis of amino acid sequence of BnSTS showed the highest 82.08% similarity with STS from Arabidopsis thaliana.The Real-Time PCR indicated that gene expression level and the concentration of stachyose in different development stage of seeds were associated.Gene expression and the acquirement of desiccation tolerance of seeds had a time consistency trend,suggesting that the gene may play an important role in seed desiccation tolerance mechanisms.