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[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。
[Objective] The PsSFBB gene of Xinjiang pear was cloned and analyzed by bioinformatics. [Method] Anther of Xinjiang Pear cultivars, Cheri Pear, was used as materials. The PsSFBB gene was cloned by RT-PCR and RACE. The cloned genes were analyzed by bioinformatics. [Result] The PsS-FBB-α gene with a total length of 1 231 bp (GenBank accession number EU909687) was cloned and named PsSFBB6-α. This gene encodes 378 amino acids and contains an F-box motif of about 50 amino acids at the N-terminus. Bioinformatics analysis showed that the molecular formula of PsSFBB6-α was C2000H3034N517O558S23, the relative molecular mass was 44 987.5 and the isoelectric point was 6.02. The secondary structure was dominated by α-helix. The theoretical half-life was about 30 h and the unstable parameters were 55.21, belong to unstable protein; and presumably this protein is hydrophilic, non-secreted protein, play a role in the matrix lyase, specific recognition substrate, just in line with the role of F-box protein. [Conclusion] This study laid the foundation for further study on the mechanism of SFBB protein and self-incompatibility, and provided theoretical basis for the scientific configuration of pollination tree in the breeding and production of Xinjiang pear self-compatibility varieties to improve the yield and quality.