Proof

Protein Modification

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Microbial Consortia

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R-R System

Introduction

R-R (Reporting and Regulation)system is constructed by us in order to report the expression of PETase gene and regulate the expression process. In this section there are two main concepts we need to prove. The first is whether the ddpX gene can cause cell lysis and another is whether the ddpX gene can cause cell lysis under the CpxR promoter when inclusion body form because of the overexpression of PETase gene. We constructed the part BBa_K2110008 to prove the validity of this concept.

Constructing Process

This part is modified and improved from the former part BBa_K339007 by changing the original mRFP gene to the novel ddpX gene (Part: BBa_K2110008). The ddpX gene was obtained by colony PCR of E.coli. Considering there is no restriction endonuclease cutting site among the subparts, we used PCR to amplify the CpxR promoter-RBS sequence and linked it with the plasmid backbone and the ddpX gene. Then the new part was obtained.

Proof Result

After we constructed the new biobrick, we firstly used PCR to amplify the whole biobrick and linked it to the recombinant plasmid pUC19 already with PETase gene in it. Then we transfromed the plasmid into E.coli and induced the expression of PETase gene. Then we measured the OD₆₀₀ of the culture medium. The result is showed below.