Part:BBa_K3612011

Pseudoalteromonas terminator (TaspC) TaspC is a terminator from Pseudoalteromonas genus, which will be used in all our genetic designs to stop the AFPs transcription.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE 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 TaspC terminator, in order to control the expression of our protein type of interest, AFP, in Pseudoalteromonas nigrifaciens. TaspC will be used to stop AFP expression when expressing in Pseudoalteromonas nigrifaciens.

Biology

TaspC is a strong rho-independent transcription terminator of the aspartate aminotransferase gene from Pseudoalteromonas haloplanktis TAC125. Used with the Pasp promoter (Part number: K3612014), result in a successful expression of alpha amylase in bacteria of Pseudoalteromonas genus (5, 6)

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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.

Secondary Structure

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Measurement

• [http://openwetware.org/wiki/Cconboy:Terminator_Characterization/Results How these parts were measured]