Part:BBa_K3612014
Pseudoalteromonas constitutive promoter (Pasp)
Pasp is a constitutive promoter from Pseudoalteromonas genus, which will be used for the AFPs expression in Pseudoalteromonas nigrifaciens.
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 Pasp promoter, in order to control the expression of our protein type of interest, AFP, in Pseudoalteromonas nigrifaciens. This way, our team will use Pasp along with its RBS (BBa_K3612029) 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
PhaspC promoter, known as Pasp, is a constitutive and cold-adapted promoter from Pseudoalteromonas haloplanktis TAC125. Used with the Tasp terminator (Part number: BBa_K3612011), result in a successful expression of alpha amylase in bacteria of Pseudoalteromonas genus (5, 6).
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) Tutino, M. L., Parrilli, E., Giaquinto, L., Duilio, A., Sannia, G., Feller, G., & Marino, G. (2002). Secretion of alpha-Amylase from Pseudoalteromonas haloplanktis TAB23: Two Different Pathways in Different Hosts. Society, 184(20), 5814–5817.
(6) Birolo L, Tutino LM, Fontanella B, Gerday C, Mainolfi K, Pascarella S, et al. Aspartate aminotransferase from the Antarctic bacterium Pseudoalteromonas haloplanktis TAC 125. Cloning, expression, properties, and molecular modelling. Eur J Biochem. 2000 May;267(9):2790–802.
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