关键词:噬菌体;干扰素;酶联免疫吸附测定;Western;blot
【摘要】 目的 为今后干扰素αlc/86D突变体文库的建立和筛选高活性的新型干扰素分子打下基础。方法 利用phage display技术表达人IFNαlc/86D基因。结果 酶联免疫吸附试验、点杂交和Western blot 均证明重组噬菌体颗粒具有干扰素的免疫反应性,生物学活性测定表明,当重组噬菌体为5×1010 TU时,与4IU的重组IFNαlb活性相当。结论 人IFNαlc/86D在噬菌体表面能正确折叠和功能性表达。
Functional phage display of human interferon alpha 1c/86D Ma Xuejun,Hu Rong,Lu Hai,et al.State Key Laboratory for Molecular Virology and Genetic Engineering,Beijing 100052
Abstract We have successfully displayed human interferon-αlc/86D(IFN-αlc/86D)on the surface of the filamentous bacteriophage using a phagemid vector syetem (pCANTAB5E).The IFN-αlc/86D cDNA was fused to a DNA sequence encoding the amino-terminal domain of pⅢ,a minor coat protein exposed at one end of the phage.Fusion gene wase packaged into phagemid particles upon superinfection with M13K07 helper phage.Expression of IFN-αlc/86D was verified by its reactivity with IFN-αl specific neutralizing antibodies and the phage displayed IFN-αlc86D was found to possess antiviral biological activity on VSV challenged WISH cells comparable to that of human recombinant IFN-αlb.These results demonstrate that IFN-αlc/86D can be displayed on phage in a correctly folded and functionally active form,and this system can be applicable to the sorting of a large repertoire of phage-displayed IFN-αlc/86D variants.
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