Cloning and expression of the rpoH gene from Escherichia coli and its product purification

被引：0

作者：

Chen, YH ^{[1
]}

Hua, ZC ^{[1
]}

Dong, C ^{[1
]}

Ma, Z ^{[1
]}

Zhu, DX ^{[1
]}

机构：

[1] NANJING UNIV, DEPT BIOCHEM, NANJING 210093, PEOPLES R CHINA

来源：

ACTA BIOCHIMICA ET BIOPHYSICA SINICA | 1996年 / 28卷 / 02期

关键词：

sigma(32); heat shock protein; gene expression; nickel-iminodiacetic acid affinity chromatography;

D O I：

暂无

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The rpoH gene, encoding the heat shock protein transcription factor sigma(32), was obtained by PCR and cloned into plasmid pUHE with the tac promoter. Upon induction with IPTG, it directed the expression of sigma(32) tailed with six additional histidine residues at its C-terminus. The histidine-tagged sigma(32) was purified with nickel-iminodiacetic acid affinity chromatography. Using S-35 radiolabeling methods in vivo, we found that even at low temperature the expression of sigma(32)-6 His may markedly increase the synthesis of the heat shock proteins such as GroEL, DnaK, and HtpH.

引用

页码：170 / 176

页数：7

共 12 条

[1] DELETION AND INSERTION MUTATIONS IN THE RPOH GENE OF ESCHERICHIA-COLI THAT PRODUCE FUNCTIONAL SIGMA-32 [J].

CALENDAR, R ;

ERICKSON, JW ;

HALLING, C ;

NOLTE, A .

JOURNAL OF BACTERIOLOGY, 1988, 170 (08) :3479-3484

[2] IS HSP70 THE CELLULAR THERMOMETER [J].

CRAIG, EA ;

GROSS, CA .

TRENDS IN BIOCHEMICAL SCIENCES, 1991, 16 (04) :135-140

[3]

HOEHULI E, 1988, J CHROMATOGR, V444, P293

[4]

HOEHULI E, 1987, J CHROMATOGR, V411, P177

[5] HISTIDINE-TAGGED RNA-POLYMERASE - DISSECTION OF THE TRANSCRIPTION CYCLE USING IMMOBILIZED ENZYME [J].

KASHLEV, M ;

MARTIN, E ;

POLYAKOV, A ;

SEVERINOV, K ;

NIKIFOROV, V ;

GOLDFARB, A .

GENE, 1993, 130 (01) :9-14

[6] A MULTIFUNCTIONAL PROKARYOTIC PROTEIN EXPRESSION SYSTEM - OVERPRODUCTION, AFFINITY PURIFICATION, AND SELECTIVE DETECTION [J].

KROLL, DJ ;

ABDELHAFIZ, HA ;

MARCELL, T ;

SIMPSON, S ;

CHEN, CY ;

GUTIERREZHARTMANN, A ;

LUSTBADER, JW ;

HOEFFLER, JP .

DNA AND CELL BIOLOGY, 1993, 12 (05) :441-453

[7] CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].

LAEMMLI, UK .

NATURE, 1970, 227 (5259) :680-+

[8] NUCLEOTIDE-SEQUENCE OF THE HEAT-SHOCK REGULATORY GENE OF ESCHERICHIA-COLI SUGGESTS ITS PROTEIN PRODUCT MAY BE A TRANSCRIPTION FACTOR [J].

LANDICK, R ;

VAUGHN, V ;

LAU, ET ;

VANBOGELEN, RA ;

ERICKSON, JW ;

NEIDHARDT, FC .

CELL, 1984, 38 (01) :175-182

[9] THE DNAK CHAPERONE MODULATES THE HEAT-SHOCK RESPONSE OF ESCHERICHIA-COLI BY BINDING TO THE SIGMA-32 TRANSCRIPTION FACTOR [J].

LIBEREK, K ;

GALITSKI, TP ;

ZYLICZ, M ;

GEORGOPOULOS, C .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (08) :3516-3520

[10]

Maniatis T., 1989, MOL CLONING

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