水稻肽链释放因子OseRF1-3基因克隆、表达及启动子分析  

Cloning,Expression and Promoter Analysis of Polypeptide Release Factor OseRF1-3 Gene from Rice

作　　者：唐跃辉 包欣欣 刘坤[1,2] 张慧聪 王双 赵君苇 娄慧敏 王箐 梁静 乔蓉 李成伟 

机构地区：[1]周口师范学院植物遗传与分子育种重点实验室,河南周口466000 [2]河南省作物分子育种与生物反应器重点实验室,河南周口466000

出　　处：《华北农学报》2018年第1期1-6,共6页Acta Agriculturae Boreali-Sinica

基　　金：2018年度河南省高等学校重点科研项目(18A180035);河南省科技开放合作项目(132106000077);河南省科学技术厅科技攻关项目(152102410074);周口师范学院青年基金(ZKNUB3201801);高层次人才基金(ZKNUC2016030)

摘　　要：为了揭示水稻中肽链释放因子eRF1在蛋白质生物合成中的作用,对水稻eRF1基因的全长cDNA和启动子进行了克隆与表达模式分析。通过RT-PCR技术克隆获得1个水稻eRF1基因,命名为OseRF1-3,序列分析表明,该基因CDS序列全长1 308 bp,编码436个氨基酸,包含N、M、C共3个保守结构域。qRT-PCR结果表明,OseRF1-3是组成型表达,在水稻穗中表达最高。以水稻叶片基因组DNA为模板,通过PCR技术克隆了该基因起始密码子上游2 120 bp启动子序列,构建该基因启动子融合GUS植物表达载体,并通过农杆菌介导法导入水稻愈伤组织。GUS组织化学染色分析表明,OseRF1-3基因启动子驱动GUS基因在水稻各组织部位都表达即组成型表达,在根中表达较弱,在水稻花、硬壳中检测到GUS基因较强表达。GUS检测OseRF1-3基因启动子驱动GUS表达的结果与qRT-PCR得出的结果相一致。因此,该研究将为进一步探索OseRF1-3基因的功能奠定理论基础。To reveal the function NA and promoter （2 120 bp） of rice ted that the OseRF1-3 contained a 1 M and C domains, qRT-PCR results of rice polypeptide release factor eRF1 in protein synthesis,the full length cD- eRF1 gene,named OseRF1-3,was isolated by PCR. Sequence analysis indica- 308 bp CDS sequence that encoded 436 indicated that the transcript of OseRF1-3 amino acids with three conserved N, was detected in all of the tissues ex- amined,but the highest level of expression was detected in spikes. The genome DNA from leaves of rice was used as the template, the 2 120 bp promoter sequence of the initiation codon upstream of OseRF1-3 gene were cloned using the PCR method. The promoter of OseRF1-3 gene was fused with GUS reporter gene and transferred into Oryza sati- va L. callus by Agrobacterium-mediated method. Histochemical staining of different organs of the transgenic plants showed that histochemical GUS expression could be found in all of the tissues examined, and a low GUS reporter gene expression in root and higher expression in flowers and rice husk. The results would provide the basis for the understanding of the OseRF1-3 gene function. The results of GUS gene expression driven by the promoter of OseRF1-3 gene were in good agreement with the results from qRT-PCR analysis. So the results will provide the ba- sis for the further explore of the OseRF1-3 gene function.

关 键 词：水稻 肽链释放因子 启动子 表达模式 GUS 

分 类 号：S511.03[农业科学—作物学]