Cloning and stable maintenance of 300-kilobase-pair fragments of human DNA in Escherichia coli using an F-factor-based vector (original) (raw)

Abstract

A bacterial cloning system for mapping and analysis of complex genomes has been developed. The BAC system (for bacterial artificial chromosome) is based on Escherichia coli and its single-copy plasmid F factor. It is capable of maintaining human genomic DNA fragments of greater than 300 kilobase pairs. Individual clones of human DNA appear to be maintained with a high degree of structural stability in the host, even after 100 generations of serial growth. Because of high cloning efficiency, easy manipulation of the cloned DNA, and stable maintenance of inserted DNA, the BAC system may facilitate construction of DNA libraries of complex genomes with fuller representation and subsequent rapid analysis of complex genomic structure.

8794

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abremski K., Hoess R., Sternberg N. Studies on the properties of P1 site-specific recombination: evidence for topologically unlinked products following recombination. Cell. 1983 Apr;32(4):1301–1311. doi: 10.1016/0092-8674(83)90311-2. [DOI] [PubMed] [Google Scholar]
  2. Bellis M., Gérard A., Charlieu J. P., Marçais B., Brun M. E., Viegas-Péquignot E., Carter D. A., Roizès G. Construction and characterization of a partial library of yeast artificial chromosomes from human chromosome 21. DNA Cell Biol. 1991 May;10(4):301–310. doi: 10.1089/dna.1991.10.301. [DOI] [PubMed] [Google Scholar]
  3. Burke D. T., Carle G. F., Olson M. V. Cloning of large segments of exogenous DNA into yeast by means of artificial chromosome vectors. Science. 1987 May 15;236(4803):806–812. doi: 10.1126/science.3033825. [DOI] [PubMed] [Google Scholar]
  4. Green E. D., Riethman H. C., Dutchik J. E., Olson M. V. Detection and characterization of chimeric yeast artificial-chromosome clones. Genomics. 1991 Nov;11(3):658–669. doi: 10.1016/0888-7543(91)90073-n. [DOI] [PubMed] [Google Scholar]
  5. Hosoda F., Nishimura S., Uchida H., Ohki M. An F factor based cloning system for large DNA fragments. Nucleic Acids Res. 1990 Jul 11;18(13):3863–3869. doi: 10.1093/nar/18.13.3863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ishiura M., Hazumi N., Koide T., Uchida T., Okada Y. A recB recC sbcB recJ host prevents recA-independent deletions in recombinant cosmid DNA propagated in Escherichia coli. J Bacteriol. 1989 Feb;171(2):1068–1074. doi: 10.1128/jb.171.2.1068-1074.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kim U. J., Shizuya H., de Jong P. J., Birren B., Simon M. I. Stable propagation of cosmid sized human DNA inserts in an F factor based vector. Nucleic Acids Res. 1992 Mar 11;20(5):1083–1085. doi: 10.1093/nar/20.5.1083. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kohara Y., Akiyama K., Isono K. The physical map of the whole E. coli chromosome: application of a new strategy for rapid analysis and sorting of a large genomic library. Cell. 1987 Jul 31;50(3):495–508. doi: 10.1016/0092-8674(87)90503-4. [DOI] [PubMed] [Google Scholar]
  9. O'Connor M., Peifer M., Bender W. Construction of large DNA segments in Escherichia coli. Science. 1989 Jun 16;244(4910):1307–1312. doi: 10.1126/science.2660262. [DOI] [PubMed] [Google Scholar]
  10. Rackwitz H. R., Zehetner G., Frischauf A. M., Lehrach H. Rapid restriction mapping of DNA cloned in lambda phage vectors. Gene. 1984 Oct;30(1-3):195–200. doi: 10.1016/0378-1119(84)90120-3. [DOI] [PubMed] [Google Scholar]
  11. Rackwitz H. R., Zehetner G., Murialdo H., Delius H., Chai J. H., Poustka A., Frischauf A., Lehrach H. Analysis of cosmids using linearization by phage lambda terminase. Gene. 1985;40(2-3):259–266. doi: 10.1016/0378-1119(85)90048-4. [DOI] [PubMed] [Google Scholar]
  12. Sternberg N. Bacteriophage P1 cloning system for the isolation, amplification, and recovery of DNA fragments as large as 100 kilobase pairs. Proc Natl Acad Sci U S A. 1990 Jan;87(1):103–107. doi: 10.1073/pnas.87.1.103. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Wada M., Little R. D., Abidi F., Porta G., Labella T., Cooper T., Della Valle G., D'Urso M., Schlessinger D. Human Xq24-Xq28: approaches to mapping with yeast artificial chromosomes. Am J Hum Genet. 1990 Jan;46(1):95–106. [PMC free article] [PubMed] [Google Scholar]
  14. Wyman A. R., Wolfe L. B., Botstein D. Propagation of some human DNA sequences in bacteriophage lambda vectors requires mutant Escherichia coli hosts. Proc Natl Acad Sci U S A. 1985 May;82(9):2880–2884. doi: 10.1073/pnas.82.9.2880. [DOI] [PMC free article] [PubMed] [Google Scholar]