栽培大豆染色体改构复合体基因SNP5的克隆及序列分析

Cloning and Sequence Analysis of a SNF5-type Chromatin-remodeling Complex Gene in Glycine max

根据大豆耐盐基因表达芯片上提供的表达量下调的EST序列信息,借助了电子克隆方法,根据获得的假定序列设计引物,通过RT-PCR,从栽培大豆克隆了染色体改构复合体SNP5类同源基因。其片段长度为812bp,编码240个氨基酸的多肽,分子重量为27.4kD,等电点pI为5.37。NCBI保守结构域分析表明,其属于SNF5超家族,因此我们将其命名为Gm-SNP5(GenBank登录号:HM068595)。Southern杂交表明,GmSNF5基因在栽培大豆基因中至少有一个拷贝。氨基酸序列及系统进化树分析表明,GmSNF5与其同科的豌豆PsSNF5有较高的同源性,氨基酸序列一致性高达92%。由于其部分EST序列在大豆耐盐芯片中表达量下调,因此推测此基因在功能上与栽培大豆的耐盐性相关。

Based on the down-regulated EST sequence information in the salt tolerant gene expression chip,a SNF5-type Chromatin-remodeling complex gene was cloned from Glycine max by RT-PCR,using primers designed according to the sequence in silico cloning.It is 812 bp in length encoded an ORF of 240 amino acids with a theoretical molecular weight of 27.4 kD and isoelectric point of 5.37.The conserved domain analysis in NCBI showed that it belongs to SNF5 superfamily,and named as GmSNP5（GenBank Accession number：HM068595）.Southern blot analysis revealed that it has at least one copy in G.max genome.Amino acid sequence and phylogenetic analysis showed that GmSNF5 is most similar to the Pisum sativum SNF5 protein with 92% identity at the amino acid level.As part of its EST sequences was down-regulated in the tolerant soybean chips,it suggests that this gene is functionally related to salt tolerance in G.max.