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PMID: 6308658 Published · ppublish English Journal Article

Replacement and amplification of bacterial genes with sequences altered in vitro.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 80 ·No. 16 ·1983-08-00 ·Pages 4894-8

Gutterson NI, Koshland DE

Abstract

An efficient method for the replacement of chromosomal DNA by segments altered in vitro has been developed for bacteria. The method requires (i) a recombinant plasmid with a ColE1-like replicon and (ii) a strain defective in DNA polymerase I (polA), which is unable to replicate the plasmid extrachromosomally. This method is of general use since there are a number of suitable vectors and polA strains are available in both Escherichia coli and Salmonella typhimurium, the two most widely studied bacterial species. Using the method, we have constructed two chromosomal deletions in the chemotaxis gene region of S. typhimurium. In addition, plasmid sequences integrated into the chromosome have been amplified up to 30-fold by varying the concentration of ampicillin or tetracycline in the growth medium.

MeSH Terms
Chromosomes, Bacterial/physiology Cloning, Molecular DNA Polymerase I/genetics DNA, Recombinant/metabolism DNA-Directed DNA Polymerase/genetics Drug Resistance, Microbial Escherichia coli/enzymology,genetics Gene Amplification Genes, Bacterial Genetic Vectors Kinetics Plasmids Salmonella/enzymology,genetics Tetracycline/pharmacology
Chemicals
DNA, Recombinant DNA Polymerase I DNA-Directed DNA Polymerase Tetracycline
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Gutterson N I
Koshland D E
References (31)
31 references, click to expand
  1. Replacement of chromosome segments with altered DNA sequences constructed in vitro.
    Proc Natl Acad Sci U S A. 1979 Oct;76(10):4951-5 PMID: 388424
  2. Rapid and efficient cosmid cloning.
    Nucleic Acids Res. 1981 Jul 10;9(13):2989-98 PMID: 6269067
  3. Biochemistry of sensing and adaptation in a simple bacterial system.
    Annu Rev Biochem. 1981;50:765-82 PMID: 6791579
  4. Separation of signal transduction and adaptation functions of the aspartate receptor in bacterial sensing.
    Science. 1983 Jun 3;220(4601):1016-20 PMID: 6302843
  5. Isolation, characterization and complementation of Salmonella typhimurium chemotaxis mutants.
    J Mol Biol. 1975 Sep 15;97(2):225-35 PMID: 1100857
  6. Quantitation of the sensory response in bacterial chemotaxis.
    Proc Natl Acad Sci U S A. 1975 Feb;72(2):710-3 PMID: 1091931
  7. Bacterial resistance to the tetracyclines.
    Microbiol Rev. 1978 Dec;42(4):707-24 PMID: 739968
  8. Transductional Heterogenotes in Escherichia Coli.
    Genetics. 1956 Sep;41(5):758-79 PMID: 17247661
  9. Molecular cloning of chemotaxis genes and overproduction of gene products in the bacterial sensing system.
    J Bacteriol. 1981 Aug;147(2):390-400 PMID: 7021528
  10. A rapid boiling method for the preparation of bacterial plasmids.
    Anal Biochem. 1981 Jun;114(1):193-7 PMID: 6269464
  11. Use of cloned mtl genes of Escherichia coli to introduce mtl deletion mutations into the chromosome.
    J Bacteriol. 1983 Feb;153(2):685-92 PMID: 6401703
  12. Detection of specific sequences among DNA fragments separated by gel electrophoresis.
    J Mol Biol. 1975 Nov 5;98(3):503-17 PMID: 1195397
  13. Selective multiplication of dihydrofolate reductase genes in methotrexate-resistant variants of cultured murine cells.
    J Biol Chem. 1978 Mar 10;253(5):1357-70 PMID: 627542
  14. Tandem genetic duplications in phage and bacteria.
    Annu Rev Microbiol. 1977;31:473-505 PMID: 334045
  15. Conditional lethality of recA and recB derivatives of a strain of Escherichia coli K-12 with a temperature-sensitive deoxyribonucleic acid polymerase I.
    J Bacteriol. 1972 Mar;109(3):971-8 PMID: 4551758
  16. Developmental cell interactions in Myxococcus xanthus and the spoC locus.
    Proc Natl Acad Sci U S A. 1983 Mar;80(5):1406-10 PMID: 16593290
  17. Transduction of linked genetic characters of the host by bacteriophage P1.
    Virology. 1955 Jul;1(2):190-206 PMID: 13267987
  18. Mechanism of integrating foreign DNA during transformation of Bacillus subtilis.
    Proc Natl Acad Sci U S A. 1978 Aug;75(8):3664-8 PMID: 99740
  19. Analysis of restriction fragments of T7 DNA and determination of molecular weights by electrophoresis in neutral and alkaline gels.
    J Mol Biol. 1977 Feb 15;110(1):119-46 PMID: 845942
  20. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system.
    Gene. 1977;2(2):95-113 PMID: 344137
  21. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid.
    J Bacteriol. 1978 Jun;134(3):1141-56 PMID: 149110
  22. Functional homology of chemotaxis genes in Escherichia coli and Salmonella typhimurium.
    J Bacteriol. 1979 Jul;139(1):107-14 PMID: 378950
  23. Genetic aspects of bacterial endospore formation.
    Bacteriol Rev. 1976 Dec;40(4):908-62 PMID: 12736
  24. Establishment of Escherichia coli cells with an integrated high copy number plasmid.
    Mol Gen Genet. 1980;178(3):525-33 PMID: 6993851
  25. Isolation of an E. coli strain with a mutation affecting DNA polymerase.
    Nature. 1969 Dec 20;224(5225):1164-6 PMID: 4902142
  26. Deletion mutants of bacteriophage lambda. I. Isolation and initial characterization.
    J Mol Biol. 1971 Mar 14;56(2):369-84 PMID: 4323931
  27. Determination of nucleic acid sequence homologies and relative concentrations by a dot hybridization procedure.
    Nucleic Acids Res. 1979 Nov 24;7(6):1541-52 PMID: 503860
  28. The origin of replication of plasmid p15A and comparative studies on the nucleotide sequences around the origin of related plasmids.
    Cell. 1983 Jan;32(1):119-29 PMID: 6186390
  29. Transformation and preservation of competent bacterial cells by freezing.
    Methods Enzymol. 1979;68:326-31 PMID: 396437
  30. Gene amplification and gene correction in somatic cells.
    Cell. 1982 May;29(1):109-19 PMID: 7105178
  31. Isolation and characterization of a mutant of Salmonella typhimurium deficient in a major deoxyribonucleic acid polymerase activity.
    J Bacteriol. 1973 Oct;116(1):54-8 PMID: 4355486
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.80.16.4894
Published
1983-08-00
Pages
4894-8
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC384153
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.80.16.4894 PMC Full Text View journal details
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