Difference between revisions of "Part:BBa K5103001"

Revision as of 02:36, 29 September 2024

PCG004 with Cry8Da

Sequence and Features

Profile

Name: PCG004 with Cry8Da

Base Pairs: 12661 bp

Origin: Add

Properties: Add

Usage and Biology

It includes features for selective gene expression in a dropout region that normally contains green fluorescent protein (GFP) flanked by BsaI sites. In this case we replaced GFP with Cry8Da crystal toxin sequences from Bacillus thuringiensis (Bt). Upstream, a lacI sequence encodes the lac repressor and a lac operator (lacO) overlaps with the pGrac promoter. Using this plasmid allows us to induce the expression of Cry8Da when we want using IPTG. This allows transcription of Cry8Da by causing a conformational change in Lac, releasing it from lacO.

This is a composite part that includes pCG004, a shuttle plasmid backbone designed for E. coli , with the Cry8Da gene inserted. The Cry8Da gene is under the control of an IPTG-inducible promoter, enabling regulation of protein expression.

This composite part has been demonstrated to work effectively in E. coli DH5α, and the expression of Cry8Da is regulated using the IPTG-inducible promoter. This system can be used for producing the Cry8Da protein in E. coli for laboratory studies, protein purification, and potential applications in bioinsecticide development.

INSERT INFORMATION ABOUT THE HISTORY OF THE PART

Figure 1 . Streak plate of PCG-Cry8Da transformant colonies grown on ampicillin selective media.

Figure 2. PCG-CRY plasmid components were developed using SnapGene.