犬瘟热病毒疫苗变异株的分离及其P1蛋白的表达、鉴定

Isolation of Canine Distemper Virus Vaccine Variant Strain and the Expression and Identification of P1 Recombinant Protein in E.coli

从河北某宠物医院病犬的脾脏中分离到1株犬瘟热病毒,H基因测序结果表明,该分离株与疫苗株的同源性为99.5%,有3个氨基酸突变,但不影响其糖基化位点。将该株病毒P1基因克隆到pET32a(+)载体,转化大肠杆菌BL21(DE3)及Rosetta感受态细胞,在IPTG诱导下获得大小为50ku,可溶性表达的P1重组蛋白。Western blotting、间接ELISA试验结果显示表达产物能被犬瘟热高免血清所识别,表明其具有良好的反应原性,有望用于犬瘟热的检测。

A canine distemper virus was isolated from an infected dog in a pet hospital in Hebei. It had 99.5% homology with vaccine strain by sequencing of H gene, and three amino acids changed that had no effect on glycosylation sites. The P1 gene of this strain was cloned into pET32a（ ＋ ） vector. The recombinant plasmid was transformed into BL21 （DE3）, Rosetta and protein expression was induced by IPTG. The expression protein had a molecular weight of 50 ku. And expressed mainly in form of dissolubility body. The expressed protein reacted specially with CDV serum in Western blotting and ELISA. IFA re- sult showed that the P1 protein fixed in the cell nucleus.