一个水稻DEAD盒RNA解旋酶基因OsBIRH1的克隆鉴定及其蛋白纯化

克隆鉴定了一个水稻DEAD盒RNA解旋酶基因OsBIRH1。OsBIRH1基因全长cDNA由2 167个核苷酸组成,包含1个长1 926个核苷酸的开放阅读框,编码1个由641个氨基酸组成的蛋白,预测分子量为70.8kD。OsBIRH1基因位于水稻第3号染色体上,由8个外显子和7个内含子组成。预测的OsBIRH1蛋白包含DEAD盒解旋酶所特有的保守DEAD和HELICc结构域以及保守模体结构。系统进化树分析表明,OsBIRH1与拟南芥和人DEAD盒解旋酶有较高的同源性,氨基酸序列同一性为47%~54%。原核表达了OsBIRH1基因,纯化获得重组OsBIRH1融合蛋白,为OsBIRH1生化与生物学功能研究提供科学依据。

Molecular characterization of a rice DEAD-box RNA helicase gene OsBIRH1 and purification of recombinant protein

A rice gene,OsBIRH1,encoding for a DEADbox RNA helicase,was cloned and characterized.The full-length cDNA of OsBIRH1 was composed of 2 167 nucleotides,in which including an open reading frame(ORF)of 1 926 nucleotides in size.The predicted translation product of this ORF contains 641 amino acids with a calculated molecular weight of 70.8 kD.The OsBIRH1 gene is located on the chromosome 3 of rice genome and consists of 8 exons and 7 introns.Conserved DEAD and HELICc domains and motifs,which are characteristics of the DEAD-box RNA helicases,were identified in OsBIRH1 protein.Phylogenetic tree analysis illustrated that OsBIRH1 had higher homology with Arabidopsis DEAD-box RNA helicase and a human DEAD-box RNA helicase,sharing 47%-54% amino acid identities.The recombinant OsBIRH1 protein was over-expressed and purified from E.coli,which will be helpful in elucidating the biochemical and biological functions of OsBIRH1 in future studies.