Part:BBa_K3612030
RBS of pSHAB
Ribosome binding site of the pSHAB promoter from Pseudoalteromonas genus.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Usage
In recent years, the marine bacteria of the genus Pseudoalteromonas has been proposed as a non-conventional expression organism for low temperatures, due to its optimal growth in cold conditions and its ability to express “easy proteins” (1). With this purpose in mind, designed expression systems have successfully produced recombinant proteins in bacteria from this genus (2-4). Consequently, our team chose elements from these expression systems previously reported, like the pSHAB promoter, in order to control the expression of our protein type of interest, AFP, in Pseudoalteromonas nigrifaciens. This way, our team will use pSHAB along with its RBS (this part) and the reporter GFP (BBa_K3612010) to characterize the promoter, and with that, choose the most fit promoter for expression of the AFPs in Pseudoalteromonas nigrifaciens.
Biology
The ribosome binding site of the inducible promoter pSHAB (Part number: K3612012) from Pseudoalteromonas haloplanktis TAC125 allowed the expression of beta galactosidase in bacteria of Pseudoalteromonas genus (3,5)
References
(1) Sannino F, Giuliani M, Salvatore U, Apuzzo GA, de Pascale D, Fani R, et al. A novel synthetic medium and expression system for subzero growth and recombinant protein production in Pseudoalteromonas haloplanktis TAC125. Appl Microbiol Biotechnol. 2017 Jan 27;101(2):725–34.
(2) Parrilli E, Tutino ML. Psychrophiles: From Biodiversity to Biotechnology: Second Edition. Psychrophiles From Biodivers to Biotechnol Second Ed. 2017;1–685.
(3) Papa R, Rippa V, Sannia G, Marino G, Duilio A. An effective cold inducible expression system developed in Pseudoalteromonas haloplanktis TAC125. J Biotechnol. 2007 Jan 1;127(2):199–210.
(4) Yu ZC, Tang BL, Zhao DL, Pang X, Qin QL, Zhou BC, et al. Development of a cold-adapted Pseudoalteromonas expression system for the Pseudoalteromonas proteins intractable for the Escherichia coli system. PLoS One. 2015;10(9):1–14.
(5) Papa, R., Glagla, S., Danchin, A. et al. Proteomic identification of a two-component regulatory system in Pseudoalteromonas haloplanktis TAC125. Extremophiles. 2006; 10,483–491.
(6) Papa, R., Rippa, V., & Duilio, A. Identification of the transcription factor responsible for l-malate-dependent regulation in the marine Antarctic bacteriumPseudoalteromonas haloplanktis TAC125. FEMS Microbiology Letters. 2009;295(2), 177–186
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