Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Weifan Liang
Date created: 2013-09-13 11:00:00
Date modified: 2015-05-08 01:08:45
Thu-E Mutation Part
| Types | DnaRegion |
| Roles | Composite engineered_region |
| Sequences | BBa_K1025003_sequence (Version 1) |
Description
iGEM mut part is a plasmid constructed by 2013 iGEM Tsinghua-E team which is used for genome level in vivo high-diversity library construction. In this vector, highly error-prone dnaQ mutant, mutD[1] was cloned downstream of araBAD promoter so that we can control the mutation intensity of the target genome by the concentration of araBAD promoter???s inducer, L-arabinose in a strict manner. And the GFP downstream mutD was to test whether the mutD can be translated. Actually, we measured the mutation increase induced by our mut part with the protocol described in our note form. To be brief, by measuring the reversion of rifampinresistance caused by mutation in genome, we accurately determined the mutation rate increase to be as big as four times compared with negative control (either bacteria without mutD or without arabinose input).Notes
The genes were cloned easily by PCR and connected to plasmid backbone by in-fusion cloning.Source
MutD comes from E.coli W3110 genome with two animo acid mutation[1]. GFP was conserved in our lab and its GenBank accession number is KF020493.1.[1]Schaaper, R. M. MECHANISMS OF MUTAGENESIS IN THE ESCHERICHIA-COLI MUTATOR MUTD5 - ROLE OF DNA MISMATCH REPAIR. Proc. Natl. Acad. Sci. U. S. A.85, 8126-8130, doi:10.1073/pnas.85.21.8126 (1988)