| 학회 | 한국화학공학회 |
| 학술대회 | 2007년 가을 (10/26 ~ 10/27, 한국과학기술원) |
| 권호 | 13권 2호, p.1682 |
| 발표분야 | 생물화공 |
| 제목 | Red recombination of linear dsDNAs at the replication fork |
| 초록 | Over the past few years, recombination-mediated in vivo genetic engineering, termed Recombineering, has been developed to provide a simple and highly efficient method for DNA modification in E. coli. Especially, a simple method known as “Red recombination” based on the interplay of phage-encoded proteins, “Exo and Beta,” allows the efficient tools for Recombineering. Despite many attempts to expand its application, lack of the information on the recombination mechanism still remains as a major hurdle for the accurate design of the recombination templates. In this study, we report the empirical evidence that linear dsDNAs are recombined through single-strand annealing at the replication fork when the target is denatured during DNA replication. Using the synthetic dsDNA containing the endogeneous homology between the two different antibiotic resistance genes, Cmr and Kmr, in addition to both terminal homologies, recombination was found to be lagging-strand biased. This strongly suggests that dsDNAs were recombined at the replication fork and consequently, help us for the error-proof design for Recombineering. |
| 저자 | 임성인, 민병은, 정규열 |
| 소속 | 포항공과대 |
| 키워드 | Red recombination; dsDNAs; replication; recombineering |
| 원문파일 | 초록 보기 |
