禽巴氏杆菌C_(48-3)株成熟黏附蛋白基因cpm39的克隆及序列分析

Cloning and Sequencing of a Mature Adhesive Protein Gene cpm39 of Avian Pasteurella multocida C_(48-3)

目的:对禽巴氏杆菌C48-3株编码成熟黏附蛋白的基因cpm39进行克隆和序列分析。方法:通过PCR从禽巴氏杆菌C48-3基因组DNA中扩增出cpm39基因,克隆到pMD18-T载体中,转化大肠杆菌DH5α,并对目的基因进行核苷酸序列测定;用Clustal X和Mega 2.1软件将测定的序列与GenBank中已登录的16种血清型巴氏杆菌株核苷酸序列进行同源性分析。结果:测序结果表明cpm39基因大小为1002 bp,与已知的16个血清型巴氏杆菌cpm39基因核苷酸序列的同源性为81.5%～100%。结论:克隆得到禽巴氏杆菌C48-3株编码成熟黏附蛋白的cpm39基因,该基因在不同血清型巴氏杆菌中具有很高的同源性,该蛋白可以作为研制预防巴氏杆菌病亚单位疫苗的候选抗原。

Objective： To clone and sequence the cpm39 gene encoding mature adhesive protein of avian Pasteurella multocida C48-3. Methods： The cpm39 gene encoding mature protein was amplified from the genomic DNA of avian P.multocida C48-3 by PCR and cloned into the pMD18-T vector. The recombinant plasmid of pMD18-cpm39 was transformed into the competent E.coli DH5α cells and then sequenced. Sequence similarity searches were performed using Clustal X and Mega 2.1 software. Results： The coding region of cpm39 gene is 1 002 bp in length. After comparing with cpm39 from 16 P.multocida serotype, it showed that nucleotide sequence homologies were 81.5% to 100%, Conclusion： The encoding mature adhesive protein of cpm39 gene was successfully cloned from strain C48-3, and multiple sequence alignment of cpm39 nucleotide sequences of different serotypes revealed high homology. Therefore, the Cpm39 protein might be a useful vaccine candidate antigen for P.multocida.