鹅多杀性巴氏杆菌OmpH的扩增与生物信息学分析

为扩增鹅多杀性巴氏杆菌OmpH基因,预测OmpH蛋白二级结构和B细胞抗原表位,通过PCR扩增测序及生物信息学分析技术对该基因序列进行开放阅读框(ORF)的寻找、编码氨基酸的推导及蛋白质二级结构的初步预测等,探讨了鹅多杀性巴氏杆菌OmpH基因在免疫保护中的作用。结果表明:鹅多杀性巴氏杆菌OmpH基因全长1537bp,ORF包含1056bp编码351个氨基酸;二级结构以无规则卷曲为主,有少量的α-螺旋和延伸带;推测OmpH蛋白有1个细胞黏附位点、3个糖基化位点。该研究为分析鹅多杀性巴氏杆菌外膜蛋白理化特性、制备单克隆抗体和设计表位疫苗等奠定了理论基础。

Amplication and Sequence Analysis of Pasteurella multocida Outer Membrane Protein (OmpH) from Goose

For amplification the OmpH gene of Pasteurella multocida and prediction its secondary structure and B-cell epitopes,PCR amplification and bioinformatics analysis were used for searching the ORF,translating the nucleotide to protein sequence,secondary structure predication and so on,which provides a theoretical basis for the study of immune mechanism and Pasteurella multocida subunit vaccine.The results indicated that Pasteurella multocida OmpH gene was 1 537 bp,ORF including 1 056 bp,encoding 351 amino acids.Using two methods,prediction of the secondary structure of Pasteurella multocida OmpH indicated that random coils were the main structural type of the flexible region in secondary structure,and contain a small amount of α-helix and extended strand.The OmpH protein was supposed contain 1 cell attachment sequence and 3 potential glycosylated sites.These results provide a theoretical basis for the further study of immune mechanisms,monoclonal antibodies preparation and epitope vaccine design.