新疆梨PsSFBB基因克隆及其生物信息学分析

[目的]对新疆梨PsSFBB基因进行克隆,并对其进行生物信息学分析。[方法]以新疆梨品种棋盘梨的花药为材料,利用RT-PCR和RACE技术对其PsSFBB基因进行克隆,并对克隆出的基因进行生物信息学分析。[结果]试验克隆到了一个全长为1 231 bp的PsS-FBB-α基因(Genbank登录号为EU909687),命名为PsSFBB6-α。该基因编码378个氨基酸,在N末端含有一个约50个氨基酸组成的F-box基序。经生物信息学分析,推测PsSFBB6-α蛋白的分子式为C2000H3034N517O558S23,相对分子质量为44 987.5,等电点为6.02,二级结构以α螺旋为主;理论推导半衰期大约为30 h,不稳定参数为55.21,属于不稳定蛋白;并且推测此蛋白为亲水性、非分泌型蛋白,在基质内起裂合酶的作用,特异性的识别底物,正好吻合了F-box蛋白的作用。[结论]为进一步研究SFBB蛋白和自交不亲和机理奠定了基础,也为新疆梨自交亲和品种的选育和生产中科学配置授粉树以提高产量和品质提供理论了依据。

Cloning and Bioinformatics Analysis of PsSFBB Gene in Xinjiang Pear

[Objective] This study aimed to clone the PsSFBB gene from Xinjiang pear for bioinformatics analysis.[Method] PsSFBB gene was cloned from anthers of Qipan pear by using RT-PCR and RACE technologies for bioinformatics analysis.[Result] A SFBB gene with a full-length of 1 231 bp was cloned and named PsSFBB6-α(Genbank accession number: EU909685).PsSFBB6-α gene encoded a protein of 378 amino acids,with an F-box motif composed of about 50 amino acids in the N-terminal.According to the bioinformatics analysis,the molecular formula of PsSFBB6-α protein was C2000H3034N517O558S23,with relative molecular mass of 43 987.5 and isoelectric point of 6.02,and the secondary structure was dominated by α-helices;theoretically,the half life period was 30 h and the instability parameter was 55.21,so PsSFBB6-α protein was an instable protein;in addition,it was predicted that PsSFBB6-α protein was a hydrophilicity and non-secretory protein with lyases activity and specifically recognized substrates,which was consistent with the function of F-box protein.[Conclusion] This study had laid the foundation for further research of SFBB proteins and the mechanism of self-incompatibility and provided a theoretical basis for breeding of self-compatible cultivars of Xinjiang pear and scientific arrangement of pollination trees in production to increase the yield and quality.