为研究猪多杀性巴氏杆菌(Pm)重组抗菌肽ABC转运体ATP结合蛋白的免疫保护性,本研究采用PCR方法扩增该蛋白基因,并将其克隆于pET-28a(+)中构建重组质粒pET-ABP,转化大肠杆菌BL21(DE3),IPTG诱导重组蛋白表达.分析表明,猪Pm抗菌肽ABC转运体ATP结合蛋白基因为696 bp,编码231 aa,不同血清型的该蛋白基因推导氨基酸序列同源性达99％以上.表达的蛋白分子量约为27 ku.动物实验表明,实验组小鼠三免后第14d其血清IgG抗体水平显著升高,明显高于对照组.对照组攻毒后3d存活率为20％,7d全部死亡;免疫组小鼠攻毒后3d存活率为80％,7d存活率为40％.该研究为进一步研制Pm的混合疫苗奠定了基础.%To explore immune protective efficacy induced by porcine Pasteurella multocida (Pm) recombinant ABC-type antimicrobial peptide transporter ATP-binding protein,the protein-coding gene was amplicatied by PCR and then cloned into pET-28a(+) to construct pET-ABP.The pET-ABP was transformed into E.coli BL21 (DE3) and the recombination protein was expressed induced by IPTG.Bioinformatics analysis of gene sequence showed that this gene was 696 bp long encoding 231 amino acids,and the molecular weigh of the encoded protein was about 27 ku.The protein sequence had above 99％ homology with different serotypes of Pm.Animal experiments showed that the IgG antibody level in serum of mice significantly increased in immuned group after third immunization and significantly higher than that in adjuvant group.The survival rate of mice in the adjuvant group was 20％ on 3 days after challenge with Pm,and all the mice was died on 7 days after challenge,but the above values of the mice in immunized group were 80％ and 40％,respectively.This study provided basic data for screening new molecular vaccines of Pm.
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