EXPRESSION IN ESCHERICHIA-COLI AND PURIFICATION OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 CAPSID PROTEIN (P24)

被引：45

作者：

EHRLICH, LS ^{[1
]}

KRAUSSLICH, HG ^{[1
]}

WIMMER, E ^{[1
]}

CARTER, CA ^{[1
]}

机构：

[1] SUNY STONY BROOK,DEPT MICROBIOL,STONY BROOK,NY 11794

来源：

AIDS RESEARCH AND HUMAN RETROVIRUSES | 1990年 / 6卷 / 10期

关键词：

D O I：

10.1089/aid.1990.6.1169

中图分类号：

R392 [医学免疫学]; Q939.91 [免疫学];

学科分类号：

100102 ;

摘要：

Capsid protein (p24;CA) of human immunodeficiency virus type 1 (HIV-1) was synthesized in Escherichia coli strain BL21 (DE3) using a plasmid encoding a truncated HIV-1 gag/pol gene. The plasmid, which contained a mutation in the frameshift region, expressed viral proteinase (PR), a pol gene product, in the gag reading frame, resulting in efficient processing of mature CA and other gag-related products. The expressed CA is soluble, recognized by monoclonal antibodies directed against HIV CA and has an N-terminal sequence identical to that of CA purified from HIV. Purification was done under mild conditions where coexpressed HIV PR retained enzymatic activity. Milligram quantities of 90% pure CA protein were obtained after chromatography on DEAE cellulose followed by facilitated aggregation of the CA in the unbound fraction. The precipitated CA was readily dissolved in low ionic strength aqueous buffer. Gel exclusion chromatography results indicated that, in solution, CA existed in oligomeric form. © 1990, Mary Ann Liebert, Inc. All rights reserved.

引用

页码：1169 / 1175

页数：7

共 27 条

[1] ISOLATION OF A T-LYMPHOTROPIC RETROVIRUS FROM A PATIENT AT RISK FOR ACQUIRED IMMUNE-DEFICIENCY SYNDROME (AIDS) [J].