Please use this identifier to cite or link to this item: http://repositorio.unicamp.br/jspui/handle/REPOSIP/726
Type: Artigo de periódico
Title: Cloning, expression and purification of a glycosylated form of the DNA-binding protein Dps from Salmonella enterica Typhimurium
Author: HANNA, Ebert Seixas
ROQUE-BARREIRA, Maria-Cristina
MENDES, Guilherme Martines Teixeira
SOARES, Sandro Gomes
BROCCHI, Marcelo
Abstract: Dps, found in many eubacterial and archaebacterial species, appears to protect cells from oxidative stress and/or nutrient-limited environment. Dps has been shown to accumulate during the stationary phase, to bind to DNA non-specifically, and to form a crystalline structure that compacts and protects the chromosome. Our previous results have indicated that Dps is glycosylated at least for a certain period of the bacterial cell physiology and this glycosylation is thought to be orchestrated by some factors not yet understood, explaining our difficulties in standardizing the Dps purification process. In the present work, the open reading frame of the dps gene, together with all the upstream regulatory elements, were cloned into a PCR cloning vector. As a result, the expression of dps was also controlled by the plasmid system introduced in the bacterial cell. The gene was then over-expressed regardless of the growth phase of the culture and a glycosylated fraction was purified to homogeneity by lectin-immobilized chromatography assay. Unlike the high level expression of Dps in Salmonella cells, less than 1% of the recombinant protein was purified by affinity chromatography using jacalin column. Sequencing and mass spectrometry data confirmed the identity of the dps gene and the protein, respectively. In spite of the low level of purification of the jacalin-binding Dps, this work shall aid further investigations into the mechanism of Dps glycosylation. (C) 2008 Elsevier Inc. All rights reserved.
Subject: recombinant Dps
glycosylation
Salmonella
Country: Estados Unidos
Editor: ACADEMIC PRESS INC ELSEVIER SCIENCE
Rights: fechado
Identifier DOI: 10.1016/j.pep.2008.01.015
Address: http://apps.isiknowledge.com/InboundService.do?Func=Frame&product=WOS&action=retrieve&SrcApp=EndNote&UT=000256514200002&Init=Yes&SrcAuth=ResearchSoft&mode=FullRecord
http://dx.doi.org/10.1016/j.pep.2008.01.015
Date Issue: 2008
Appears in Collections:FCM - Artigos e Materiais de Revistas Científicas

Files in This Item:
File Description SizeFormat 
art_HANNA_Cloning_expression_and_purification_of_a_glycosylated_2008.pdfpublished version416.85 kBAdobe PDFView/Open


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.