Part:BBa_K3861022
hilA transcriptional regulator
hilA, together with hilC and hilD, are global regulators and control the expression of the Salmonella pathogenicity island I (SPI-1) genes in S. typhimurium on which the SPI-1 T3SS injectisome is encoded.1 Over 25 genes that are needed for host invasion are located on the SPI-1.2 Also, hilA activates the SPI-1 operon mediated by the prg and inv promoters.3,4 Thus, hilA controls SPI-1 genes in response to enviromental needs like pH and oxygen levels.4–6
As a part of our project “TargteTaxi”, we require the SPI-1 T3SS to be present in the minicells membrane. For assembly of the SPI-1 T3SS in the minicells, we must overexpress the SPI-1 encoded genes as it has been shown previously by Carleton et al. (2013)7. The overexpression of the T3SS genes can be achieved by overexpressing its master regulator, hilA. In our bacterial strain, hilA expression is going to be regulated by the arabinose inducible promoter of Salmonella Typhimurium, allowing us to control the expression of the SPI-1 encoded genes.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 1112
Illegal PstI site found at 1309 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 1112
Illegal PstI site found at 1309 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1635
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 1112
Illegal PstI site found at 1309 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 1112
Illegal PstI site found at 1309
Illegal NgoMIV site found at 151
Illegal AgeI site found at 1258 - 1000COMPATIBLE WITH RFC[1000]
References
1. Widmaier, D. M. et al. Engineering the Salmonella type III secretion system to export spider silk monomers. Mol. Syst. Biol. 5, 309 (2009).
2. Heran Darwin, K. & Miller, V. L. InvF Is Required for Expression of Genes Encoding Proteins Secreted by the SPI1 Type III Secretion Apparatus in Salmonella typhimurium. J. Bacteriol. 181, 4949–4954 (1999).
3. Bajaj, V., Hwang, C. & Lee, C. A. hilA is a novel ompR/toxR family member that activates the expression of Salmonella typhimurium invasion genes. Mol. Microbiol. 18, 715–727 (1995).
4. Bajaj, V., Lucas, R. L., Hwang, C. & Lee, C. A. Co-ordinate regulation of Salmonella typhimurium invasion genes by environmental and regulatory factors is mediated by control of hilA expression. Mol. Microbiol. 22, 703–714 (1996).
5. Ahmer, B. M., van Reeuwijk, J., Watson, P. R., Wallis, T. S. & Heffron, F. Salmonella SirA is a global regulator of genes mediating enteropathogenesis. Mol. Microbiol. 31, 971–982 (1999).
6. Main-Hester, K. L., Colpitts, K. M., Thomas, G. A., Fang, F. C. & Libby, S. J. Coordinate regulation of Salmonella pathogenicity island 1 (SPI1) and SPI4 in Salmonella enterica serovar Typhimurium. Infect. Immun. 76, 1024–1035 (2008).
7. Carleton, H. A., Lara-Tejero, M., Liu, X. & Galán, J. E. Engineering the type III secretion system in non-replicating bacterial minicells for antigen delivery. Nat. Commun. (2013) doi:10.1038/ncomms2594.
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