This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Chen, C Y
Right arrow Articles by Winans, S C
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Chen, C Y
Right arrow Articles by Winans, S C

 Previous Article  |  Next Article 

J Bacteriol. 1991 February; 173(3): 1139-1144

research-article

Controlled expression of the transcriptional activator gene virG in Agrobacterium tumefaciens by using the Escherichia coli lac promoter.

C Y Chen and S C Winans

Section of Microbiology, Cornell University, Ithaca, New York 14853.

ABSTRACT

The Agrobacterium VirG protein is normally expressed from two promoters in response to multiple stimuli, including plant-released phenolics (at promoter P1) and acidic growth media (at promoter P2). To simplify the analysis of vir gene induction, we sought to create Agrobacterium strains in which virG could be expressed in a controllable fashion. To study the possibility of using the lac promoter and repressor, we constructed a plasmid containing the lac promoter fused to the lacZ structural gene. A derivative of this plasmid containing the lacIq gene was also constructed. The plasmid not containing lacIq expressed high levels of beta-galactosidase. The plasmid containing lacIq expressed beta-galactosidase at very low levels in the absence of o-nitrophenyl-beta-D-galactoside (IPTG) and at moderate levels in the presence of IPTG. We also fused the lac promoter to a virG::lacZ translational fusion and found that IPTG elevated expression of this translational fusion to moderate levels, though not to levels as high as from the stronger of the two native virG promoters. Finally, the lac promoter was used to express the native virG gene in strains containing a virB::lacZ translational fusion. virB expression in this strain depended on addition of IPTG as well as the vir gene inducer acetosyringone. In a similar strain lacking lacIq, virB expression was greater than in a strain in which virG was expressed from its native promoters. Expression of virG from the lac promoter did not alter the acidic pH optimum for vir gene induction, indicating that the previously observed requirement for acidic media was not due solely to the need to induce P2.

J Bacteriol. 1991 February; 173(3): 1139-1144




This article has been cited by other articles:

  • Khan, S. R., Gaines, J., Roop, R. M. II, Farrand, S. K. (2008). Broad-Host-Range Expression Vectors with Tightly Regulated Promoters and Their Use To Examine the Influence of TraR and TraM Expression on Ti Plasmid Quorum Sensing. Appl. Environ. Microbiol. 74: 5053-5062 [Abstract] [Full Text]  
  • Guo, M., Jin, S., Sun, D., Hew, C. L., Pan, S. Q. (2007). Recruitment of conjugative DNA transfer substrate to Agrobacterium type IV secretion apparatus. Proc. Natl. Acad. Sci. USA 104: 20019-20024 [Abstract] [Full Text]  
  • Glenn, S. A., Gurich, N., Feeney, M. A., Gonzalez, J. E. (2007). The ExpR/Sin Quorum-Sensing System Controls Succinoglycan Production in Sinorhizobium meliloti. J. Bacteriol. 189: 7077-7088 [Abstract] [Full Text]  
  • Jakubowski, S. J., Cascales, E., Krishnamoorthy, V., Christie, P. J. (2005). Agrobacterium tumefaciens VirB9, an Outer-Membrane-Associated Component of a Type IV Secretion System, Regulates Substrate Selection and T-Pilus Biogenesis. J. Bacteriol. 187: 3486-3495 [Abstract] [Full Text]  
  • Gao, R., Lynn, D. G. (2005). Environmental pH Sensing: Resolving the VirA/VirG Two-Component System Inputs for Agrobacterium Pathogenesis. J. Bacteriol. 187: 2182-2189 [Abstract] [Full Text]  
  • Zhu, J., Chai, Y., Zhong, Z., Li, S., Winans, S. C. (2003). Agrobacterium Bioassay Strain for Ultrasensitive Detection of N-Acylhomoserine Lactone-Type Quorum-Sensing Molecules: Detection of Autoinducers in Mesorhizobium huakuii. Appl. Environ. Microbiol. 69: 6949-6953 [Abstract] [Full Text]  
  • Jakubowski, S. J., Krishnamoorthy, V., Christie, P. J. (2003). Agrobacterium tumefaciens VirB6 Protein Participates in Formation of VirB7 and VirB9 Complexes Required for Type IV Secretion. J. Bacteriol. 185: 2867-2878 [Abstract] [Full Text]  
  • Ding, Z., Christie, P. J. (2003). Agrobacterium tumefaciens Twin-Arginine-Dependent Translocation Is Important for Virulence, Flagellation, and Chemotaxis but Not Type IV Secretion. J. Bacteriol. 185: 760-771 [Abstract] [Full Text]  
  • He, X., Chang, W., Pierce, D. L., Seib, L. O., Wagner, J., Fuqua, C. (2003). Quorum Sensing in Rhizobium sp. Strain NGR234 Regulates Conjugal Transfer (tra) Gene Expression and Influences Growth Rate. J. Bacteriol. 185: 809-822 [Abstract] [Full Text]  
  • Li, L., Jia, Y., Hou, Q., Charles, T. C., Nester, E. W., Pan, S. Q. (2002). A global pH sensor: Agrobacterium sensor protein ChvG regulates acid-inducible genes on its two chromosomes and Ti plasmid. Proc. Natl. Acad. Sci. USA 99: 12369-12374 [Abstract] [Full Text]  
  • Pellock, B. J., Teplitski, M., Boinay, R. P., Bauer, W. D., Walker, G. C. (2002). A LuxR Homolog Controls Production of Symbiotically Active Extracellular Polysaccharide II by Sinorhizobium meliloti. J. Bacteriol. 184: 5067-5076 [Abstract] [Full Text]  
  • Akakura, R., Winans, S. C. (2002). Mutations in the occQ Operator That Decrease OccR-induced DNA Bending Do Not Cause Constitutive Promoter Activity. J. Biol. Chem. 277: 15773-15780 [Abstract] [Full Text]  
  • York, G. M., Stubbe, J., Sinskey, A. J. (2002). The Ralstonia eutropha PhaR Protein Couples Synthesis of the PhaP Phasin to the Presence of Polyhydroxybutyrate in Cells and Promotes Polyhydroxybutyrate Production. J. Bacteriol. 184: 59-66 [Abstract] [Full Text]  
  • Sagulenko, E., Sagulenko, V., Chen, J., Christie, P. J. (2001). Role of Agrobacterium VirB11 ATPase in T-Pilus Assembly and Substrate Selection. J. Bacteriol. 183: 5813-5825 [Abstract] [Full Text]  
  • York, G. M., Junker, B. H., Stubbe, J., Sinskey, A. J. (2001). Accumulation of the PhaP Phasin of Ralstonia eutropha Is Dependent on Production of Polyhydroxybutyrate in Cells. J. Bacteriol. 183: 4217-4226 [Abstract] [Full Text]  
  • Zhao, Z., Sagulenko, E., Ding, Z., Christie, P. J. (2001). Activities of virE1 and the VirE1 Secretion Chaperone in Export of the Multifunctional VirE2 Effector via an Agrobacterium Type IV Secretion Pathway. J. Bacteriol. 183: 3855-3865 [Abstract] [Full Text]  
  • Sagulenko, V., Sagulenko, E., Jakubowski, S., Spudich, E., Christie, P. J. (2001). VirB7 Lipoprotein Is Exocellular and Associates with the Agrobacterium tumefaciens T Pilus. J. Bacteriol. 183: 3642-3651 [Abstract] [Full Text]  
  • Peng, W.-T., Banta, L. M., Charles, T. C., Nester, E. W. (2001). The chvH Locus of Agrobacterium Encodes a Homologue of an Elongation Factor Involved in Protein Synthesis. J. Bacteriol. 183: 36-45 [Abstract] [Full Text]  
  • Rashkova, S., Zhou, X.-R., Chen, J., Christie, P. J. (2000). Self-Assembly of the Agrobacterium tumefaciens VirB11 Traffic ATPase. J. Bacteriol. 182: 4137-4145 [Abstract] [Full Text]  
  • Llosa, M., Zupan, J., Baron, C., Zambryski, P. (2000). The N- and C-Terminal Portions of the Agrobacterium VirB1 Protein Independently Enhance Tumorigenesis. J. Bacteriol. 182: 3437-3445 [Abstract] [Full Text]  
  • Prieto, M. A., Kellerhals, M. B., Bozzato, G. B., Radnovic, D., Witholt, B., Kessler, B. (1999). Engineering of Stable Recombinant Bacteria for Production of Chiral Medium-Chain-Length Poly-3-Hydroxyalkanoates. Appl. Environ. Microbiol. 65: 3265-3271 [Abstract] [Full Text]  
  • Willis, L. B., Walker, G. C. (1999). A Novel Sinorhizobium meliloti Operon Encodes an alpha -Glucosidase and a Periplasmic-Binding-Protein-Dependent Transport System for alpha -Glucosides. J. Bacteriol. 181: 4176-4184 [Abstract] [Full Text]  
  • Zhou, X.-R., Christie, P. J. (1999). Mutagenesis of the Agrobacterium VirE2 Single-Stranded DNA-Binding Protein Identifies Regions Required for Self-Association and Interaction with VirE1 and a Permissive Site for Hybrid Protein Construction. J. Bacteriol. 181: 4342-4352 [Abstract] [Full Text]  
  • Zhu, J., Winans, S. C. (1999). Autoinducer binding by the quorum-sensing regulator TraR increases affinity for target promoters in vitro and decreases TraR turnover rates in whole cells. Proc. Natl. Acad. Sci. USA 96: 4832-4837 [Abstract] [Full Text]  
  • Cheng, H.-P., Walker, G. C. (1998). Succinoglycan Is Required for Initiation and Elongation of Infection Threads during Nodulation of Alfalfa by Rhizobium meliloti. J. Bacteriol. 180: 5183-5191 [Abstract] [Full Text]  
  • Stahl, L. E., Jacobs, A., Binns, A. N. (1998). The Conjugal Intermediate of Plasmid RSF1010 Inhibits Agrobacterium tumefaciens Virulence and VirB-Dependent Export of VirE2. J. Bacteriol. 180: 3933-3939 [Abstract] [Full Text]  
  • Swiderska, A., Berndtson, A. K., Cha, M.-R., Li, L., Beaudoin, G. M. J. III, Zhu, J., Fuqua, C. (2001). Inhibition of the Agrobacterium tumefaciens TraR Quorum-sensing Regulator. INTERACTIONS WITH THE TraM ANTI-ACTIVATOR. J. Biol. Chem. 276: 49449-49458 [Abstract] [Full Text]  
  • Akakura, R., Winans, S. C. (2002). Constitutive Mutations of the OccR Regulatory Protein Affect DNA Bending in Response to Metabolites Released from Plant Tumors. J. Biol. Chem. 277: 5866-5874 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»