一株貉源犬瘟热病毒核衣壳蛋白的原核表达

Prokaryotic Expression of Nucleocapsid Protein Gene of Canine Distemper Virus from Racoon Dog

根据GenBank中犬瘟热病毒(CDV)核衣壳蛋白(N)基因的核苷酸序列设计合成1对引物,经RT-PCR从病毒总RNA中扩增出约1600bp的片段,将此片段克隆于pMD18-Tsimple载体。经测序分析表明,与其它毒株N基因序列的同源性很高。再将其定向克隆于原核表达载体pPROEXTMHTa中,转化感受态E.coli BL21细胞,经IPTG诱导表达分子量约63Ku的重组N蛋白。

A pair of pr/mers was designed according to the Genebank sequence of nucleocapsid gene of canine distemper virus, and N gene was amplified by RT-PCR and cloned into PMD18-T simple plasmid. A segment of 1 600 bp was obtained. Sequencing analysis indicates that the homology between this strain and the others is very high. Then the N gene was subcloned into Prokaryotic expression plasmid pPROEXTMHTa. Recombinant plasmid carrying N gene （pPROEXTM HT-N） was transformed into E. coli BL21 （ DE3 ） and induced with IPTG. A fusion protein about 63 Ku was expressed.