DNA cloning by homologous recombination in Escherichia coli - PubMed (original) (raw)
. 2000 Dec;18(12):1314-7.
doi: 10.1038/82449.
Affiliations
- PMID: 11101815
- DOI: 10.1038/82449
DNA cloning by homologous recombination in Escherichia coli
Y Zhang et al. Nat Biotechnol. 2000 Dec.
Abstract
The cloning of foreign DNA in Escherichia coli episomes is a cornerstone of molecular biology. The pioneering work in the early 1970s, using DNA ligases to paste DNA into episomal vectors, is still the most widely used approach. Here we describe a different principle, using ET recombination, for directed cloning and subcloning, which offers a variety of advantages. Most prominently, a chosen DNA region can be cloned from a complex mixture without prior isolation. Hence cloning by ET recombination resembles PCR in that both involve the amplification of a DNA region between two chosen points. We apply the strategy to subclone chosen DNA regions from several target molecules resident in E. coli hosts, and to clone chosen DNA regions from genomic DNA preparations. Here we analyze basic aspects of the approach and present several examples that illustrate its simplicity, flexibility, and remarkable efficiency.
Similar articles
- A short GC-rich sequence involved in deletion formation of cloned DNA in E. coli.
Sümegi A, Birkó Z, Szeszák F, Vitális S, Biró S. Sümegi A, et al. Acta Biol Hung. 1997;48(3):275-9. Acta Biol Hung. 1997. PMID: 9406607 - A new logic for DNA engineering using recombination in Escherichia coli.
Zhang Y, Buchholz F, Muyrers JP, Stewart AF. Zhang Y, et al. Nat Genet. 1998 Oct;20(2):123-8. doi: 10.1038/2417. Nat Genet. 1998. PMID: 9771703 - High-throughput cloning of human liver complete open reading frames using homologous recombination in Escherichia coli.
Zhu D, Zhong X, Tan R, Chen L, Huang G, Li J, Sun X, Xu L, Chen J, Ou Y, Zhang T, Yuan D, Zhang Z, Shu W, Ma L. Zhu D, et al. Anal Biochem. 2010 Feb 15;397(2):162-7. doi: 10.1016/j.ab.2009.10.018. Epub 2009 Oct 14. Anal Biochem. 2010. PMID: 19835833 - Ordered deletions using exonuclease III.
Clark D, Henikoff S. Clark D, et al. Methods Mol Biol. 1994;31:47-55. doi: 10.1385/0-89603-258-2:47. Methods Mol Biol. 1994. PMID: 7921037 Review. No abstract available. - [Red/ET recombination and its biomedical applications].
Wang JP, Zhang YM. Wang JP, et al. Sheng Wu Gong Cheng Xue Bao. 2005 May;21(3):502-6. Sheng Wu Gong Cheng Xue Bao. 2005. PMID: 16108384 Review. Chinese.
Cited by
- Redox Regulation of LAT Enhances T Cell-Mediated Inflammation.
James J, Coelho A, Lahore GF, Hernandez CM, Forster F, Malissen B, Holmdahl R. James J, et al. Antioxidants (Basel). 2024 Apr 22;13(4):499. doi: 10.3390/antiox13040499. Antioxidants (Basel). 2024. PMID: 38671946 Free PMC article. - Automated high-throughput DNA synthesis and assembly.
Ma Y, Zhang Z, Jia B, Yuan Y. Ma Y, et al. Heliyon. 2024 Feb 25;10(6):e26967. doi: 10.1016/j.heliyon.2024.e26967. eCollection 2024 Mar 30. Heliyon. 2024. PMID: 38500977 Free PMC article. Review. - Precise genome engineering in Pseudomonas using phage-encoded homologous recombination and the Cascade-Cas3 system.
Zheng W, Xia Y, Wang X, Gao S, Zhou D, Ravichandran V, Jiang C, Tu Q, Yin Y, Zhang Y, Fu J, Li R, Yin J. Zheng W, et al. Nat Protoc. 2023 Sep;18(9):2642-2670. doi: 10.1038/s41596-023-00856-1. Epub 2023 Aug 25. Nat Protoc. 2023. PMID: 37626246 Review. - Review of knockout technology approaches in bacterial drug resistance research.
Tong C, Liang Y, Zhang Z, Wang S, Zheng X, Liu Q, Song B. Tong C, et al. PeerJ. 2023 Aug 17;11:e15790. doi: 10.7717/peerj.15790. eCollection 2023. PeerJ. 2023. PMID: 37605748 Free PMC article. Review. - Applications of Programmable Endonucleases in Sequence- and Ligation-Independent Seamless DNA Assembly.
Xiong X, Lu Z, Ma L, Zhai C. Xiong X, et al. Biomolecules. 2023 Jun 21;13(7):1022. doi: 10.3390/biom13071022. Biomolecules. 2023. PMID: 37509059 Free PMC article. Review.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources