OVERPRODUCTION AND SINGLE-STEP PURIFICATION OF GAL4 FUSION PROTEINS FROM ESCHERICHIA-COLI

被引：44

作者：

REECE, RJ ^{[1
]}

RICKLES, RJ ^{[1
]}

PTASHNE, M ^{[1
]}

机构：

[1] HARVARD UNIV,DEPT BIOCHEM & MOLEC BIOL,CAMBRIDGE,MA 02138

来源：

GENE | 1993年 / 126卷 / 01期

关键词：

TRANSCRIPTION ACTIVATION; RECOMBINANT DNA; YEAST;

D O I：

10.1016/0378-1119(93)90596-U

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

A DNA fragment encoding the yeast GAL4 transcriptional activator DNA-binding domain (amino acids 1 93) was cloned into an Escherichia coli expression vector such that the overproduced protein is tagged with six histidine residues and a factor Xa protease cleavage site. The vector also contains unique restriction sites at the 3' end of the gene to allow the construction of fusion proteins. These fusion proteins can easily be purified to homogeneity and their activity tested in vitro.

引用

页码：105 / 107

页数：3

共 8 条

[1] AN AMINO-TERMINAL FRAGMENT OF GAL4 BINDS DNA AS A DIMER [J].

CAREY, M ;

KAKIDANI, H ;

LEATHERWOOD, J ;

MOSTASHARI, F ;

PTASHNE, M .

JOURNAL OF MOLECULAR BIOLOGY, 1989, 209 (03) :423-432

[2] REGULATION OF DNA-REPLICATION INVITRO BY THE TRANSCRIPTIONAL ACTIVATION DOMAIN OF GAL4-VP16 [J].

CHENG, LZ ;

WORKMAN, JL ;

KINGSTON, RE ;

KELLY, TJ .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (02) :589-593

[3] DNA RECOGNITION BY GAL4 - STRUCTURE OF A PROTEIN DNA COMPLEX [J].

MARMORSTEIN, R ;

CAREY, M ;

PTASHNE, M ;

HARRISON, SC .

NATURE, 1992, 356 (6368) :408-414

[4] HOW EUKARYOTIC TRANSCRIPTIONAL ACTIVATORS WORK [J].

PTASHNE, M .

NATURE, 1988, 335 (6192) :683-689

[5] THE C-TERMINAL DOMAIN OF THE ESCHERICHIA-COLI DNA GYRASE-A SUBUNIT IS A DNA-BINDING PROTEIN [J].

REECE, RJ ;

MAXWELL, A .

NUCLEIC ACIDS RESEARCH, 1991, 19 (07) :1399-1405

[6] TRANSCRIPTIONAL REPRESSION BY YY1, A HUMAN GLI-KRUPPEL-RELATED PROTEIN, AND RELIEF OF REPRESSION BY ADENOVIRUS E1A PROTEIN [J].

SHI, Y ;

SETO, E ;

CHANG, LS ;

SHENK, T .

CELL, 1991, 67 (02) :377-388

[7] MULTICOPY EXPRESSION VECTORS CARRYING THE LAC REPRESSOR GENE FOR REGULATED HIGH-LEVEL EXPRESSION OF GENES IN ESCHERICHIA-COLI [J].

STARK, MJR .

GENE, 1987, 51 (2-3) :255-267

[8] BOTH OCT-1 AND OCT-2A CONTAIN DOMAINS WHICH CAN ACTIVATE THE UBIQUITOUSLY EXPRESSED U2 SNRNA GENES [J].

YANG, J ;

MULLERIMMERGLUCK, MM ;

SEIPEL, K ;

JANSON, L ;

WESTIN, G ;

SCHAFFNER, W ;

PETTERSSON, U .

EMBO JOURNAL, 1991, 10 (08) :2291-2296

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