铁蛋白与犬α干扰素的融合表达及抗病毒活性检测  

Prokaryotic Expression and Antiviral Activity of Canine Interferon Fused with Fe Protein

作　　者：宋若楠 刘春草 李航 王真真[1,2] 贾楠楠 朱杰 李传峰[2] 程松[3] 梁留存 刘光清 王金泉[1] 孟春春[1,2] SONG Ruonan;LIU Chuncao;LI Hang;WANG Zhenzhen;JIA Nannan;ZHU Jie;LI Chuanfeng;CHENG Song;LIANG Liucun;LIU Guangqing;WANG Jinquan;MENG Chunchun(Xinjiang Agricultural University,Urumqi 830052,China;Shanghai Veterinary Research Institute,CAAS,Shanghai 200241,China;Laboklin Laboratory for Clinical Diagnostics,Shanghai 200241,China)

机构地区：[1]新疆农业大学,乌鲁木齐830000 [2]中国农业科学院上海兽医研究所,上海200241 [3]德国纳博科临中国检测实验室,上海200241

出　　处：《中国动物传染病学报》2024年第4期91-96,共6页Chinese Journal of Animal Infectious Diseases

基　　金：中国农业科学院青年英才培育工程专项经费。

摘　　要：为获得治疗犬病毒性传染病的特效药物,本研究将铁蛋白与犬α干扰素进行融合表达。首先根据GenBank中公布的犬α干扰素序列(IFN2a)合成模板,通过融合PCR连入铁蛋白编码基因的上游,并将该重组基因克隆至原核表达载体pCold-TF中,获得重组质粒pCold-TF-CaIFN2a-Fe。转化BL21后用异丙基-β-D硫代半乳糖苷(IPTG)诱导,经聚丙烯酰胺凝胶电泳(SDSPAGE),蛋白免疫印迹(Western blot)鉴定,获得约89 kDa的可溶性重组目的蛋白。重组蛋白经镍柱纯化后,使用犬肾细胞/水疱疹性口炎病毒(MDCK/VSV)微量细胞病变抑制法检测抗病毒活性,效价确定为8×104 IU/mg,显示出良好的应用前景。In this study,ferritin was fused with canine interferonαto obtain specific drugs for canine viral infectious diseases.Firstly,according to the synthesis template of canine interferonαsequence(IFN2a)published in GenBank,the ferritin encoding gene was linked to the upstream by fusion PCR and the recombinant gene was cloned into the prokaryotic expression vector pCold-TF to obtain the recombinant plasmid pCold-TF-CaIFN2a-Fe.After transformation into BL21,the recombinant protein was induced by IPTG and identified by SDS-PAGE and Western blot.The soluble recombinant protein of 89 kDa was obtained and purified with nickel column.Its antiviral activity was detected using MDCK/VSV microcytopathy inhibition assay.The antiviral titer was 8×104 IU/mg,showing a good application prospect.

关 键 词：铁蛋白 犬α干扰素 融合表达 抗病毒活性 

分 类 号：S859.79[农业科学—临床兽医学]