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作 者:张丽伟 茹毅 郝荣增 马坤 李亚军 伍春平 张姣燕 齐萌[1] 郑海学[2] ZHANG Li-wei;RU Yi;HAO Rong-zeng;MA Kun;LI Ya-jun;WU Chun-ping;ZHANG Jiao-yan;QI Meng;ZHENG Hai-xue(College of Animal Science,Tarim University,Aral 843300,China;State Key Laboratory of Veterinary Etiological Biology/National Foot-and-Mouth Disease Reference Lob oratory/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China)
机构地区:[1]塔里木大学动物科学学院,新疆阿拉尔843300 [2]中国农业科学院,兰州兽医研究所,家畜疫病病原生物学国家重点实验室,国家口蹄疫参考实验室,甘肃兰州730046
出 处:《中国兽医科学》2021年第10期1233-1238,共6页Chinese Veterinary Science
基 金:国家自然科学基金项目(31672585)。
摘 要:为了研究重组猪干扰素α6(rPoIFN-α6)对口蹄疫病毒(FMDV)的抑制作用,建立rPoIFN-α6的CHO细胞分泌表达工艺,随后采用亲和层析纯化获得rPoIFN-α6,该rPoIFN-α6的生物活性为1.7×10^(7)U/mg。细胞病变抑制结果显示,以10^(5)倍倍比稀释的rPoIFN-α6预处理PK-15细胞,可以完全抑制100 TCID_(50)的A/FMDV的增殖;在1×10^(4)TCID_(50)的A/FMDV感染后的48 h,rPoIFN-α6也可以完全抑制A/FMDV的增殖。该稀释度的rPoIFN-α6也可以完全抑制1×10^(3)TCID_(50)的O/FMDV的增殖,且在O/FMDV感染后的32 h,仍然可以完全抑制O/FMDV的增殖。结果表明,获得的rPoIFN-α6可有效抑制A/GDMM/CHA/2013和O/GDBY/CHA/2010两种FMDV毒株的增殖,为新型基因工程抗病毒制剂的开发以及口蹄疫的预防和治疗奠定了基础。In order to study the inhibitory effect of recombinant porcine interferon α6(rPoIFN-α6)on foot-and-mouth disease virus(FMDV),the secretory expression process of rPoIFN-α6 in CHO cells was established,and then affinity chromatography was used to purify the obtained rPoIFN-α6.The biological activity of it was 1.7×10^(7) U/mg.The results of cytopathic inhibition showed that PK-15 cells pretreated with rPoIFN-α6 diluted by a factor of 10^(5) can effectively inhibit the proliferation of 100 TCID_(50) A/FMDV.During the 48 hours after infection with 1×10^(4) TCID_(50) A/FMDV,rPoIFN-α6 can still effectively constrain the proliferation of A/FMDV.rPoIFN-α6 of the same dilution can also inhibit the proliferation of O/FMDV with 1 ×10^(3) TCID_(50) to a large degree,and it can continously restrain the proliferation of O/FMDV in the following 32 hours after the O/FMDV infection.The results show that the obtained rPoIFN-α6 can successfully inhibit the proliferation of two FMDV strains,A/GDMM/CHA/2013 and O/GDBY/CHA/2010.This study is not only useful for the development of new genetically engineered antiviral agents but also helpful in the prevention and treatment of foot-and-mouth disease.
关 键 词:猪干扰素α6 CHO悬浮细胞 表达与纯化 口蹄疫病毒
分 类 号:S852.659.6[农业科学—基础兽医学]
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