---

作 者 ：张雨良;王希东;张桦;杨峰山;姚正培;石庆华*

期 刊 ：植物研究 2009年 29卷 4期 页码：445-452

关键词：大麦；HvYrg1基因；克隆；RNA干扰；

Keywords:Hordeum vulgare, HvYrg1 gene, cloning, RNA interfere,

摘 要 ：大麦谷粒是受多个数量性状基因(QTL)控制的复杂性状，而RING E3泛素连接酶在决定大麦产量和蛋白质降解途径中起到极为重要的作用。本研究按照同源克隆的方法，依据水稻、拟南芥、玉米、小麦和酵母等E3泛素连接酶保守区域设计引物，采用RT-PCR方法从西藏大麦中克隆出产量相关基因HvYrg1全长cDNA序列，包括完整的开放阅读框架(ORF)1 275 bp，编码蛋白为424个氨基酸(GenBank No. EU333863)。同源性比较结果显示，它与GenBank上已报道的水稻GW2基因同源性最高为86%。以植物表达载体pCAMBIA2300-35s-OCS质粒为基础，构建由35s启动子调控的HvYrg1基因的RNA干扰载体pCAM-RNAi-HvYrg1。这一载体的成功构建为研究该基因在作物产量的功能鉴定打下了很好的基础。

Abstract:Barley grain is a complex trait controlled by QTL, and RING E3 ubiquitin ligase plays a vital role in grain yield and protein degradation pathway. According to the conservative region of E3 ubiquitin ligase family homologous sequences, such as rice, Arabidopsis, corn, wheat and yeast, one pair of specific primers was designed and used to clone the corresponding gene from barley by RTPCR amplification. The results showed that a full-length cDNA sequence of HvYrg1 gene from Tibetan barley was obtained, which includes a integrity of open reading frame (ORF) 1 275 bp, encoding 424 amino acids (GenBank No. EU333863). Homology comparison showed that HvYrg1 gene shared 86% identity with rice GW2 gene reported in GenBank. Based on plant expression vector pCAMBIA2300-35s-OCS plasmid, we constructed a plant RNAi vector driven by 35 s promoter, namely pCAM-RNAi-HvYrg1 to further investigate its functions on crop yield formation.

---