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Active transcription promotes single-stranded oligonucleotide mediated gene repair
- Source :
- Biochemical and biophysical research communications. 353(1)
- Publication Year :
- 2006
-
Abstract
- The lambda-Red-mediated recombination has been exploited as an efficient means for DNA manipulation. We previously reported that replication plays a pivotal role during this process. Replication direction dictates strand bias, such that single stranded oligonucleotide (SSO) with sequence corresponding to the nascent lagging strand directs higher levels of recombinant formation compared to its complementary SSO. In addition, the Escherichia coli methyl-directed mismatch repair system impedes efficient SSO-mediated site-specific gene repair. However, the role of transcription in determining strand bias and recombination efficiency is unclear. To address the potential role of transcriptional processes, we constructed plasmid substrates that harbor a mutant antibiotic reporter under the control of an inducible promoter. We found that transcription activation can promote recombinant formation to more than 10-folds whilst it has negligible effect on strand bias. Our findings provide evidence for a role of transcription in SSO-mediated gene repair process.
- Subjects :
- DNA, Bacterial
Recombination, Genetic
Transcriptional Activation
DNA Repair
DNA repair
Targeted Gene Repair
Biophysics
Cell Biology
Biology
Biochemistry
Molecular biology
chemistry.chemical_compound
Sense strand
chemistry
Transcription (biology)
Coding strand
Escherichia coli
DNA mismatch repair
Molecular Biology
Gene
DNA
DNA Damage
Subjects
Details
- ISSN :
- 0006291X
- Volume :
- 353
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Biochemical and biophysical research communications
- Accession number :
- edsair.doi.dedup.....60ee76098989a90e0e0d6603101afd32