中文摘要：

为更好研究猪瘟病毒（Classical swine fever virus,CSFV）N^pro蛋白的生物学功能,RT-PCR扩增CSFV的N^pro基因,克隆至p Cold I质粒并转化BL21（DE3）表达菌。经IPTG诱导,成功表达了约24 k Da的重组蛋白His-N^pro,用镍离子亲和层析柱进行纯化后,制备了兔源多克隆抗体。Western blot结果显示,制备的多克隆抗体能特异性识别猪瘟病毒感染PK15细胞内表达的约23k Da的N^pro蛋白,但此抗体不能用于N^pro蛋白的间接免疫荧光（indirect immunofluorescence assay,IFA）检测。纯化的重组蛋白His-N^pro和制备的N^pro多抗为N^pro蛋白功能研究奠定了基础。

英文摘要：

To better understand the biological function of N^pro protein of Classical swine fever virus（CSFV）,N^pro gene amplified in RTPCR was inserted into pCold I plasmid and then transformed into E.coli BL21（DE3）.Following induction with IPTG,approx 24 kDa recombinant protein His-N^pro was expressed and purified using Ni-NTZ resin.Afterwards,specific polyclonal antibodies were prepared by immunizing rabbit with purified His-N^pro protein.The N^pro protein was detected in CSFV-infected PK15 cells in Western blot,but not in indirect immunofluorescence assay.In conclusion,the availability of recombinant protein His-N^pro and polyclonal anti-N^pro antibodies has established foundation for further study of N^pro biological function.