Difference between revisions of "Part:BBa K2243011"

Latest revision as of 19:04, 1 November 2017

TP901-1 attL_J23119_TP901-1 attR

The constitutive promoter J23119 is between TP901-1 RDF specific sites and will be inverted by the TP901-1 recombinase and its RDF(or their fusion protein), thus leading to the direction of promoter and change of gene expression. This unit can be used as standard reporter of recombinase TP901-1-RDF fusion protein characterization

Usage

We constructed this part to characterize the recombination efficiency of the excisionase Bxb1 gp35-gp47 fusion protein (BBa_K2243013). It consists of a constitutive promoter (BBa_J23119) flanked by attL and attR sites of recombinase Bxb1 gp35. Upon recombination, the orientation of the constitutive promoter change. As a result, expression of downstream sequence is shut down, and upstream sequence is transcribed.

Biology

J23119 is the consensus sequence of a combinatorial constitutive promoter library family. In nature, attL and attR is generated when Mycobacteriophage Bxb1 integrates its DNA at the attB site of the Mycobacterium smegmatis genome using the viral attP site. With RDF Bxb1 gp47, attL and attR can be recognized and undergo recombination. We obtained the promoter, attL and attR sites by oligo synthesis.

Figure 1. The standard testing system used to characterize the recombinases.

Design note

This part is constructed by Gibson Assembly.

Characterize

We plan to use this part to characterize the recombination efficiency of the integrase Bxb1 gp35 (BBa_K2243013). However, the characterization have not done yet.

Sequence and Features