Expression and purification of a putative H-NS nucleoid-associated protein from the phytopathogen Xylella fastidiosa

被引：3

作者：

Paula, DP

Azzoni, AR

Siqueira, SF

Catani, CF

Rosselli, LK

de Souza, AP

机构：

[1] Univ Estadual Campinas, Inst Biol, Dept Genet & Evolut, Genet Engn & Mol Biol, BR-13083970 Campinas, SP, Brazil

[2] Univ Estadual Campinas, Inst Biol, Dept Microbiol & Immunol, Campinas, SP, Brazil

来源：

PROTEIN EXPRESSION AND PURIFICATION | 2003年 / 32卷 / 01期

关键词：

H-NS protein; Xylella fastidiosa; DNA-binding activity or functional genomic;

D O I：

10.1016/S1046-5928(03)00193-1

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The H-NS protein is one of the major constituents of the nucleoid structure that has been implicated in the DNA packaging and in the global regulation of gene expression. The study of this transcriptional regulator is an effort to fight Xylella fastidiosa, a citrus pathogen responsible for a range of economically important plant diseases, including the citrus variegated chlorosis (CVC). The putative H-NS ORF was cloned into a pET32-Xa/LIC vector in order to overexpress it coupled with fusion tags in Escherichia Coli BL21(DE3). The expressed recombinant protein was purified by immobilized metal affinity chromatography (Ni-NTA resin) and its identity verified by mass spectrometry (MALDI-TOF). Final purification was performed by cation-exchange chromatography (SP Sepharose Fast Flow) and the purified protein was found as a single band on SDS-PAGE. The folding and its DNA binding activity were verified by circular dichroism and fluorescence spectroscopy, respectively. (C) 2003 Elsevier Inc. All rights reserved.

引用

页码：61 / 67

页数：7

共 50 条

[31] The putative Orf4 of broad-host-range conjugative plasmid R446 could be related to the H-NS family of bacterial nucleoid-associated proteins
Nieto, JM
Juárez, A
PLASMID, 1999, 41 (02) : 125 - 127
[32] DNA-Binding Properties of YbaB, a Putative Nucleoid-Associated Protein From Caulobacter crescentus
Pal, Parul
Modi, Malvika
Ravichandran, Shashank
Yennamalli, Ragothaman M.
Priyadarshini, Richa
FRONTIERS IN MICROBIOLOGY, 2021, 12
[33] Mycobacterium tuberculosis nucleoid-associated DNA-binding protein H-NS binds with high-affinity to the Holliday junction and inhibits strand exchange promoted by RecA protein
Sharadamma, N.
Harshavardhana, Y.
Singh, Pawan
Muniyappa, K.
NUCLEIC ACIDS RESEARCH, 2010, 38 (11) : 3555 - 3569
[34] THE CHROMATIN-ASSOCIATED PROTEIN H-NS
USSERY, DW
HINTON, JCD
JORDI, BJAM
GRANUM, PE
SEIRAFI, A
STEPHEN, RJ
TUPPER, AE
BERRIDGE, G
SIDEBOTHAM, JM
HIGGINS, CF
BIOCHIMIE, 1994, 76 (10-11) : 968 - 980
[35] The nucleoid-associated protein StpA binds curved DNA, has a greater DNA-binding affinity than H-NS and is present in significant levels in hns mutants
Sonnenfield, JM
Burns, CM
Higgins, CF
Hinton, JCD
BIOCHIMIE, 2001, 83 (02) : 243 - 249
[36] SYNTHESIS OF THE ESCHERICHIA-COLI K-12 NUCLEOID-ASSOCIATED DNA-BINDING PROTEIN H-NS IS SUBJECTED TO GROWTH-PHASE CONTROL AND AUTOREGULATION
DERSCH, P
SCHMIDT, K
BREMER, E
MOLECULAR MICROBIOLOGY, 1993, 8 (05) : 875 - 889
[37] EXPRESSION OF THE GENE ENCODING THE MAJOR BACTERIAL NUCLEOID PROTEIN H-NS IS SUBJECT TO TRANSCRIPTIONAL AUTO-REPRESSION
FALCONI, M
HIGGINS, NP
SPURIO, R
PON, CL
GUALERZI, CO
MOLECULAR MICROBIOLOGY, 1993, 10 (02) : 273 - 282
[38] H-NS Regulates Gene Expression and Compacts the Nucleoid: Insights from Single-Molecule Experiments
Winardhi, Ricksen S.
Yan, Jie
Kenney, Linda J.
BIOPHYSICAL JOURNAL, 2015, 109 (07) : 1321 - 1329
[39] AN UNUSUAL SUICIDAL INTERACTION IN ESCHERICHIA-COLI INVOLVING NUCLEOID PROTEIN H-NS
JAYASHREE, P
GOWRISHANKAR, J
JOURNAL OF GENETICS, 1995, 74 (1-2) : 1 - 17
[40] EXPRESSION OF THE ESCHERICHIA-COLI DIMORPHIC GLUTAMIC-ACID DECARBOXYLASES IS REGULATED BY THE NUCLEOID PROTEIN H-NS
YOSHIDA, T
YAMASHINO, T
UEGUCHI, C
MIZUNO, T
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 1993, 57 (09) : 1568 - 1569

← 1 2 3 4 5 →