人新型干扰素κ的基因克隆、表达、纯化及其抗病毒活性的初步研究  

Cloning, expression and purification of interferon-κ,a novel human interferon, and its antiviral activity

作　　者：张辉[1] 段招军[1] 朱建高[1] 彭夫望[1] 李武平[1] 高寒春[1] 谢志萍[1] 王玉娥[1] 侯云德[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所病毒基因工程国家重点实验室,北京100052

出　　处：《中华实验和临床病毒学杂志》2005年第3期223-226,共4页Chinese Journal of Experimental and Clinical Virology

摘　　要：目的制备人干扰素κ(hIFNκ)并初步研究其生物学活性。方法克隆hIFNκ的cDNA并根据大肠埃希菌的密码子偏好性人工合成了hIFNκ基因,在大肠埃希菌DH5α中高效表达、纯化后测定了rhIFNκ的多种生物学活性,包括抗病毒活性、抗细胞增殖活性、种属特异性以及NK细胞调节活性。结果成功地获得了高纯度的rhIFNκ,纯度在90%以上。利用WISHVSV系统检测rhIFNκ的抗病毒活力为2.0×106IU/mg。与rhIFNα2b的比较发现,rhIFNκ无论在抗病毒活性、细胞生长抑制,还是在促NK细胞活性方面均弱于前者。结论rhIFNκ在不同种属来源细胞上的抗病毒活性因种属来源而异,不同的病毒对rhIFNκ的敏感性是不同的。Objective To prepare human interferon-k （hIFN-k） and study its biological activities. Methods Whole length of hIFN-k＇s cDNA was cloned, and its sequence was chemically synthesized according to the optimized codons of E. coli, then was expressed in E. coli DH5α After purified, the rhIFN- k protein was tested for its various kinds of biological activities. ResuIts The purity of rhIFN-k was above 90%. In WHIS-VSV system, the antiviral activity of rhIFN-k was 2.0 × 10^6 IU/mg. Compared with rhIFN- α2b, the biological activities of rhIFN-k were all feebler, including antiviral activity, promoting NK cell activity and anti-proliteration activity. Conclusion Antiviral activities of rhIFN-k on cell lines of different species are different, different viruses show different sensitivitv to rhIFN-k.

关 键 词：干扰素类 抗病毒药 抗病毒活性 基因克隆 人干扰素 步研究 纯化 NK细胞活性 生物学活性 大肠埃希菌 

分 类 号：R346[医药卫生—基础医学]