Difference between revisions of "Part:BBa K1638035"

Revision as of 14:18, 11 September 2015

hTrx-based scaffold fused to T18 through intein and a flexible linker

This composite part consists of the hTrx-scaffold (used for presenting peptide aptamers) fused to the T18 domain of CyaA (used in the bacterial two-hybrid system) through intein and a linker. This part is used in the bacterial two-hybrid screening for peptide aptamers against specific targets. When a target candidate is fused to the T25 domain of CyaA, and a peptide aptamer binds the target, the T18 and T25 domain is brought into proximity of one another and the formation of cAMP is induced. When combined with a cAMP reporter system, like a system promoting the transcription of RFP, the presence of red-flourescent bacteria indicates an interaction between the peptide aptamer and the target protein.

Intein is added

The Library

In oder to use this part for peptide aptamer screening, a random nucleotide library has to be inserted into the scaffold. This library codes for a random peptide library that enables screening of a diverse set of peptide aptamers with different specificity. Typically, the library consist of 20 sets of the codon NNK (where N is A,T,G or C and K is G or C). See design for further notes on design of library.

Sequence and Features