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以植物Rho家族成员osRACD为诱饵 ,采用酵母双杂交体系从水稻幼穗中分离到肌动蛋白基因家族的一个新成员 Act.借助 5′RACE技术克隆到该基因的cDNA全长 ,其中含有一个1 1 34bp的读码框 ,编码 377个氨基酸和一个终止密码子 .同源性比较显示与其他已知的水稻肌动蛋白有 81 .8%~ 96 %的相似性 .生物信息学方法分析了蛋白的修饰位点、基因的结构和进化关系 .Southern杂交显示Act是单拷贝基因 ,RT PCR显示该基因在多种组织有广泛的表达 ,但在不同发育阶段表达量有明显变化 .还就Rho家族和肌动蛋白家族在进化和功能上的关系进行了讨论
Using the plant Rho family member osRACD as bait, a new member Act of the Actin gene family was isolated from young rice by yeast two-hybrid system. The full-length cDNA of this gene was cloned by 5’RACE technology and contains a 1 A 34 bp open reading frame encoding a protein of 377 amino acids and a stop codon.The homology comparison showed 81.8% ~ 96% similarity with other known rice actin.Analysis of protein , The structure and evolution of the gene.Southern hybridization shows Act is a single copy of the gene, RT PCR showed that the gene has a wide range of expression in a variety of tissues, but at different developmental stages of significant changes in expression.Also Rho family The relationship between actin family evolution and function was discussed