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。

关键词: 水稻, 肽链释放因子, 启动子, 表达模式, GUS

Abstract: To reveal the function of rice polypeptide release factor eRF1 in protein synthesis,the full length cDNA and promoter (2 120 bp) of rice eRF1 gene,named OseRF1-3,was isolated by PCR.Sequence analysis indicated that the OseRF1-3 contained a 1 308 bp CDS sequence that encoded 436 amino acids with three conserved N,M and C domains.qRT-PCR results indicated that the transcript of OseRF1-3 was detected in all of the tissues examined,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 sativa 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 basis for the further explore of the OseRF1-3 gene function.

Key words: Rice, Polypeptide release factor, Promoter, Expression pattern, GUS

中图分类号: 

• S511.03

• Q78