大豆Usp1基因的克隆和表达分析

Cloning and Expression Analysis of Usp1 Gene from Soybean

在对大豆根系盐胁迫抑制差减杂交文库EST序列分析的基础上,利用RT-PCR技术克隆得到了一个大豆Us-pA基因,命名为GmUsp1,其编码一个包含164个氨基酸残基的多肽链。多序列比对和编码蛋白结构分析表明,GmUsp1属于泛应激蛋白(Universal Stress Protein)家族成员之一。其氨基酸序列与蚕豆Vf_enod18序列相似度最高,属于MJ-0577类中含有ATP结合位点的UspA亚族,与MJ-0577有相似的蛋白二级结构。分别采用250 mmol.L-1NaCl、100μmol.L-1ABA和30%PEG 6000进行胁迫处理,耐盐品种文丰7号和盐敏感品种Union的GmUsp1均被诱导表达,二者对胁迫诱导响应时间和表达量存在差异,说明GmUsp1可能参与大豆对非生物逆境胁迫的应答调控。

According to the EST sequences of the Suppression Subtractive Hybridization（SSH）cDNA libraries,a soybean UspA gene named by GmUspl was cloned from the soybean by RT-PCR.The complete cDNA of GmUsp1 was 1 047 bp,it encoded a peptide which contained 164 amino acid residues.The 495 bp CDS showed that it belonged to the universal stress protein（UspA）family,by using the BLASTp tool of NCBI database.Homology analysis showed GmUsp1 had the highest similarity with the bean Vf_enod18,and it belonged to the group of UspA,which represented by the ATP-binging structure of the MJ-0577 protein from Methanococcus jannaschii.It also had the similar protein secondary structure with MJ-0577.In the salt sensitive variety Union and tolerant variety Wenfeng No.7,the expression of GmUsp1 was induced by 250 mmol·L-1 NaCl,100 μmol·L-1 ABA and 30% PEG-600 treatment,however,it had different response time and expression level.So GmUsp1 might be involved in plant stress adaptation,it suggested that GmUsp1 may play a role in salt stress adaptation in soybean.