Over-expression, purification, and confirmation of Bacillus anthracis transcriptional regulator NprR.
Quorum sensing (QS) has been recognized as an important biological phenomenon in which bacterial cells communicate and coordinate their gene expression and cellular processes with respect to population density. Bacillus anthracis is the etiological agent of fatal pulmonary anthrax infections, and the NprR/NprX QS system may be involved in its pathogenesis. NprR, renamed as aqsR for anthrax quorum sensing Regulator, is a transcriptional regulator that may control the expression of genes required for proliferation and survival. Currently, there is no protocol reported to over-express and purify B. anthracis AqsR. In this study, we describe cloning, purification, and confirmation of functional full-length B. anthracis AqsR protein. The AqsR gene was cloned into the pQE-30 vector with an HRV 3C protease recognition site between AqsR and the N-terminal His6-tag in order to yield near native AqsR after the His-tag cleavage, leaving only two additional amino acid residues at the N-terminus.
Center for Pharmaceutical Biotechnology, University of Illinois at Chicago, 900 S. Ashland, IL 60607, USA; Department of Medicinal Chemistry and Pharmacognosy, University of Illinois at Chicago, 900 S. Ashland, IL 60607, USA.
Amy J
AJ
Rice
Center for Pharmaceutical Biotechnology, University of Illinois at Chicago, 900 S. Ashland, IL 60607, USA.
Jerry K
JK
Woo
Center for Pharmaceutical Biotechnology, University of Illinois at Chicago, 900 S. Ashland, IL 60607, USA.
Attiya
A
Khan
Center for Pharmaceutical Biotechnology, University of Illinois at Chicago, 900 S. Ashland, IL 60607, USA.
Michael Z
MZ
Szypulinski
Center for Pharmaceutical Biotechnology, University of Illinois at Chicago, 900 S. Ashland, IL 60607, USA; Department of Medicinal Chemistry and Pharmacognosy, University of Illinois at Chicago, 900 S. Ashland, IL 60607, USA.
Michael E
ME
Johnson
Center for Pharmaceutical Biotechnology, University of Illinois at Chicago, 900 S. Ashland, IL 60607, USA; Department of Biopharmaceutical Sciences, University of Illinois at Chicago, 900 S. Ashland, IL 60607, USA. Electronic address: hlee31@uic.edu.
Hyunwoo
H
Lee
Center for Pharmaceutical Biotechnology, University of Illinois at Chicago, 900 S. Ashland, IL 60607, USA; Department of Medicinal Chemistry and Pharmacognosy, University of Illinois at Chicago, 900 S. Ashland, IL 60607, USA. Electronic address: danielhl@uic.edu.
Hyun
H
Lee
D001408:Bacillus anthracis / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D002460:Cell Line; D015870:Gene Expression; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D053038:Quorum Sensing; D020349:Surface Plasmon Resonance; D053719:Tandem Mass Spectrometry; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
D001426:Bacterial Proteins; D014157:Transcription Factors
Bacillus anthracis; Purification; Quorum sensing; The neutral protease regulator (NprR)