油菜BnTIP2;3的克隆及生物信息学分析

Cloning and Bioinformatics Analysis of BnTIP2;3 from Brassica napus

为研究水孔蛋白分子功能,采用RT-PCR技术从油菜(Brassica napus L.)‘双11’中克隆得到1个液泡膜内在蛋白(tonoplast intrinsic proteins,TIPs)类的水孔蛋白基因,命名为BnTIP2;3,并对其进行生物信息学分析。采用ProtParam、SignalP5.0、TMHMM 2.0、CDD、ProtScale、SOPMA、SWISS-MODEL等生物学软件对该蛋白质的理化性质、信号肽、跨膜结构域、保守结构域、亲水性、二级和三级结构进行预测分析。结果表明,BnTIP2;3由250个氨基酸组成,无信号肽、含有6个跨膜区以及2个NPA保守结构域,属于稳定的疏水性蛋白质。该蛋白二级结构主要由无规则卷曲、α-螺旋、β-折叠及β-转角组成,三级结构与二级结构结果一致。系统进化分析表明,BnTIP2;3与拟南芥、亚麻芥、台湾南芥亲缘关系较近。成功克隆了BnTIP2;3基因,为进一步探讨BnTIP2;3基因的生物学功能奠定了基础。

To investigate the molecular function of aquaporin,a tonoplast intrinsic proteins(TIPs)gene TIP was isolated from Brassica napus L.’zhong shuang 11’by RT-PCR,and named as BnTIP2;3.The study performed a series of bioinformatics analysis by online biological software,such as ProtParam,SignalP 5.0,TMHMM 2.0,CDD,ProtScale,SOPMA and SWISS-MODEL to analyze the physicochemical properties,transmembrane domain,signal peptide,hydrophilicity,secondary and tertiary structure.The results showed that SlTIP open reading frame had a length of 753 bp,encoding 250 amimo acid residues without peptide,furthermore,it included 6 transmembrane domains and 2 NPA(Asn-Pro-Ala)motif.The secondary structure of BnTIP2;3 protein was mainly composed of randon coil,α-helix,β-turn and extended strand.Phylogenetic analysis displayed that BnTIP2;3 protein is grouped with Arabidopsis,Camelina sativa,Arabis alpine.Here the study successfully isolated BnTIP2;3 gene in Brassica napus,which lays a foundation for further exploring the biologic function of BnTIP2;3 gene.