禽多杀性巴氏杆菌PlpB基因交叉保护力研究

CLONING THE PLPB GENE OF PASTEURELLA MULTOCIDA AND BIOLOGICAL CHARACTERIZATION OF THE EXPRESSING PROTEINS

根据报道的PM70菌株的PlpB基因序列,设计PlpB基因一对寡核苷酸引物。以C48-1菌株为实验材料,成功扩增PlpB基因的全序列,该片段共831bp。将扩增基因插入原核表达载体pGEX-KG,成功构建了重组表达质粒pKG-PlpB,并对pKG-PlpB重组表达质粒进行序列测定,结果表明:扩增序列与GenBank上所报道的PM70株的PlpB基因序列核苷酸同源性在99%以上。将该重组质粒转化大肠杆菌BL21(DE3)感受态细胞,经诱导获得大小约56KDa的表达蛋白。Western blot检测表明,该表达蛋白具有良好的反应原性。

The oligonucleotide primers for amplifying the PlpB gene of PMC48-1 strain were designed according to the sequence of gene of PM70 strain. The amplified gene which is 831bp was inserted into the prokaryote expression vector pGEX-KG, and the resulted recombinant plasmid pKG-PlpB was sequenced. This sequence processes a homology of more than 99% compared with the published sequence of PlpB gene of PM70 in GenBank. The pKG-PIpB was transformed into E.eoli BL21 （DE3） competent cells. After induction by IPTG, a 56 kDa recombinant protein was expressed. Western blot analysis revealed that the recombinant protein have a good reaction with antisera of fowl cholera.