Difference between revisions of "Part:BBa K2921400"

Revision as of 10:14, 18 October 2019

Strong Promoter + RBS + MBP + Double Terminator

This construct constitutively expresses a colored metal-binding protein MBP (Basic part: BBa_K346004) MBP is a lead binding peptide that can accumulate lead ions from aquatic environment.

Construct Design

We inserted an upstream constitutive strong promoter-RBS combination (Basic part: BBa_K880005) and a downstream strong double terminator (Basic part: BBa_B0015) so as to maximize plasmid and protein expression.

PCR

PCR check results, using the primers VF2 and VR, and sequencing results by Tri-I Biotech confirm this construct. (insert photo)

Characterization

To verify MBP expression in E. coli, we ran a SDS-PAGE gel, expecting a protein band at 17.48 kDa. We prepared and lysed overnight liquid cultures of cells expressing MBP-mRFP, MBP only, and mRFP only. The SDS-PAGE reveals a darker band at 17.48 kDa as compared to the MBP-only and mRFP-only controls. (insert photo)

Sequence and Features