Difference between revisions of "Part:BBa K1621005"
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This part contains the coding sequence of a bacterioferritin monomer derived from ''Treponema pallidum'', TpF1. It was isolated from the subspecies ''pallidum'' which causes syphilis, a disease that is transmittable by sexual contact or congenitally. Worldwide, more than 12 million new cases of syphilis are reported which underlines the need of high detection methods that are highly sensitive and specific as well as easy to perform. | This part contains the coding sequence of a bacterioferritin monomer derived from ''Treponema pallidum'', TpF1. It was isolated from the subspecies ''pallidum'' which causes syphilis, a disease that is transmittable by sexual contact or congenitally. Worldwide, more than 12 million new cases of syphilis are reported which underlines the need of high detection methods that are highly sensitive and specific as well as easy to perform. | ||
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Revision as of 20:09, 29 August 2015
TpF1 - bacterioferritin derived from Treponema pallidum
This part contains the coding sequence of a bacterioferritin monomer derived from Treponema pallidum, TpF1. It was isolated from the subspecies pallidum which causes syphilis, a disease that is transmittable by sexual contact or congenitally. Worldwide, more than 12 million new cases of syphilis are reported which underlines the need of high detection methods that are highly sensitive and specific as well as easy to perform.
TpF1 was shown to be highly immunogenic in humans and rabbits (McGill et al., 2010), what was the reason for Jiang et al. (2013) to use it for the establishment of a diagnostic ELISA for clinical applications.
Bacterioferritins in general are central players in bacterial iron metabolism. Their functions reach from detoxification of iron in the cell to iron storage and even protection against oxidative stress (Carrondo 2003). Disuflide bonds and hydrophobic interactions mediate the formation of homo-24-mers of about 190 kDa in the outer membrane (Radolf et al. 1987). Treatment with β-mercaptoethanol or boiling in SDS leads to dissaggregation of the complex, resulting in smaller oligomers (~160 kDa) and monomers (~19 kDa). This part’s sequence encodes for such a monomer (Jiang et al. 2013).
The sequence was synthesized by Integrated DNA Technologies in a variant that was codon optimized for expression in Escherichia coli with the respective tool from IDT. It was shipped to the registry in standard pSB1C3 and begins with a start codon (ATG). Cloning into the shipping backbone was performed by Gibson Assembly.