|
|||||
|
|
| 噬菌体表面表达人干扰素αlc/86D | |
| 引用本文： | 马学军,胡荣,吕海. 噬菌体表面表达人干扰素αlc/86D[J]. 中华实验和临床病毒学杂志, 1998, 12(3): 213-216 |
| 作者姓名： | 马学军 胡荣 吕海 |
| 作者单位： | 中国预防医学科学院病毒学研究所病毒基因工程国家重点实验室，大连医科大学，第一军医大学南方医院 |
| 基金项目： | 国家863高科技发展计划资助 |
| 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ｐｈａｇｅｄｉｓｐｌａｙ技术表达人ＩＦＮαｌｃ／８６Ｄ基因。结果酶联免疫吸附试验、点杂交和Ｗｅｓｔｅｒｎｂｌｏｔ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５×１０１０ＴＵ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。 |
| 关 键 词： | 噬菌体 干扰素 酶联免疫吸附测定 Westernblot |
Functional phage display of human interferon alpha 1c/86D |
|
| Ma Xuejun,Hu Rong,Lu Hai,et al.. Functional phage display of human interferon alpha 1c/86D[J]. Chinese journal of experimental and clinical virology, 1998, 12(3): 213-216 | |
| Authors: | Ma Xuejun Hu Rong Lu Hai et al. |
| Affiliation: | State Key Laboratory for Molecular Virology and Genetic Engineering, Beijing 100052. |
| Abstract: | We have successfully displayed human interferon-alpha lc/86D(IFN-alpha lc/86D) on the surface of the filamentous bacteriophage using a phagemid vector system (pCANTAB5E). The IFN-alpha lc/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 was packaged into phagemid particles upon superinfection with M13K07 helper phage. Expression of IFN-alpha lc/86D was verified by its reactivity with IFN-alpha l specific neutralizing antibodies and the phage displayed IFN-alpha lc86D was found to possess antiviral biological activity on VSV challenged WISH cells comparable to that of human recombinant IFN-alpha lb. These results demonstrate that IFN-alpha lc/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-alpha lc/86D variants. |
| Keywords: | Phage display Interferon Western blot Enzyme-linked immunosorbent assay. |
| 本文献已被 CNKI 万方数据 等数据库收录！ | |