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. 1986 Apr;52(1):101–109. doi: 10.1128/iai.52.1.101-109.1986

In vivo repackaging of recombinant cosmid molecules for analyses of Salmonella typhimurium, Streptococcus mutans, and mycobacterial genomic libraries.

W R Jacobs, J F Barrett, J E Clark-Curtiss, R Curtiss 3rd
PMCID: PMC262204  PMID: 2937735

Abstract

Strains of Escherichia coli K-12 were constructed that permitted the amplification of in vitro-packaged recombinant cosmid-transducing particles by in vivo repackaging of recombinant cosmid molecules. Thermal induction of these thermoinducible, excision-defective lysogens containing recombinant cosmid molecules yielded high titers of packaged recombinant cosmids and low levels of PFU. These strains were used to amplify packaged recombinant cosmid libraries of Mycobacterium leprae, Mycobacterium vaccae, Salmonella typhimurium, and Streptococcus mutans DNA. Contiguous and noncontiguous libraries were compared for the successful identification of cloned genes. Construction of noncontiguous libraries allowed the dissociation of desired genes from genes that were deleterious to the survival of a cosmid recombinant and permitted selection for unlinked traits that resulted in a selected phenotype. In vivo repackaging of recombinant cosmids permitted amplification of the original in vitro-packaged collection of transducing particles, storage of cosmid libraries as phage lysates, facilitation of complementation screening, expression analysis of repackaged recombinant cosmids after UV-irradiated cells were infected, in situ enzyme or immunological screening, and facilitation of recovery of recombinant cosmid molecules containing transposon inserts.

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Selected References

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  1. Bedbrook J. R., Ausubel F. M. Recombination between bacterial plasmids leading to the formation of plasmid multimers. Cell. 1976 Dec;9(4 Pt 2):707–716. doi: 10.1016/0092-8674(76)90134-3. [DOI] [PubMed] [Google Scholar]
  2. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  4. Bukhari A. I., Zipser D. Mutants of Escherichia coli with a defect in the degradation of nonsense fragments. Nat New Biol. 1973 Jun 20;243(129):238–241. doi: 10.1038/newbio243238a0. [DOI] [PubMed] [Google Scholar]
  5. CLARK A. J., MARGULIES A. D. ISOLATION AND CHARACTERIZATION OF RECOMBINATION-DEFICIENT MUTANTS OF ESCHERICHIA COLI K12. Proc Natl Acad Sci U S A. 1965 Feb;53:451–459. doi: 10.1073/pnas.53.2.451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. CURTIS S. R., 3rd CHROMOSOMAL ABERRATIONS ASSOCIATED WITH MUTATIONS TO BACTERIOPHAGE RESISTANCE IN ESCHERICHIA COLI. J Bacteriol. 1965 Jan;89:28–40. doi: 10.1128/jb.89.1.28-40.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Clark-Curtiss J. E., Curtiss R., 3rd Analysis of recombinant DNA using Escherichia coli minicells. Methods Enzymol. 1983;101:347–362. doi: 10.1016/0076-6879(83)01026-5. [DOI] [PubMed] [Google Scholar]
  8. Clark-Curtiss J. E., Jacobs W. R., Docherty M. A., Ritchie L. R., Curtiss R., 3rd Molecular analysis of DNA and construction of genomic libraries of Mycobacterium leprae. J Bacteriol. 1985 Mar;161(3):1093–1102. doi: 10.1128/jb.161.3.1093-1102.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Clarke L., Carbon J. Biochemical construction and selection of hybrid plasmids containing specific segments of the Escherichia coli genome. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4361–4365. doi: 10.1073/pnas.72.11.4361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Collins J., Hohn B. Cosmids: a type of plasmid gene-cloning vector that is packageable in vitro in bacteriophage lambda heads. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4242–4246. doi: 10.1073/pnas.75.9.4242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Curtiss R., 3rd, Charamella L. J., Stallions D. R., Mays J. A. Parental functions during conjugation in Escherichia coli K-12. Bacteriol Rev. 1968 Dec;32(4 Pt 1):320–348. [PMC free article] [PubMed] [Google Scholar]
  12. Enquist L. W., Skalka A. Replication of bacteriophage lambda DNA dependent on the function of host and viral genes. I. Interaction of red, gam and rec. J Mol Biol. 1973 Apr 5;75(2):185–212. doi: 10.1016/0022-2836(73)90016-8. [DOI] [PubMed] [Google Scholar]
  13. Erlich H. A., Cohen S. N., McDevitt H. O. A sensitive radioimmunoassay for detecting products translated from cloned DNA fragments. Cell. 1978 Apr;13(4):681–689. doi: 10.1016/0092-8674(78)90218-0. [DOI] [PubMed] [Google Scholar]
  14. Feiss M., Margulies T. On maturation of the bacteriophage lambda chromosome. Mol Gen Genet. 1973 Dec 31;127(4):285–295. doi: 10.1007/BF00267099. [DOI] [PubMed] [Google Scholar]
  15. Feiss M., Siegele D. A., Rudolph C. F., Frackman S. Cosmid DNA packaging in vivo. Gene. 1982 Feb;17(2):123–130. doi: 10.1016/0378-1119(82)90064-6. [DOI] [PubMed] [Google Scholar]
  16. Frey J., Bagdasarian M., Feiss D., Franklin F. C., Deshusses J. Stable cosmid vectors that enable the introduction of cloned fragments into a wide range of gram-negative bacteria. Gene. 1983 Oct;24(2-3):299–308. doi: 10.1016/0378-1119(83)90090-2. [DOI] [PubMed] [Google Scholar]
  17. Friedman A. M., Long S. R., Brown S. E., Buikema W. J., Ausubel F. M. Construction of a broad host range cosmid cloning vector and its use in the genetic analysis of Rhizobium mutants. Gene. 1982 Jun;18(3):289–296. doi: 10.1016/0378-1119(82)90167-6. [DOI] [PubMed] [Google Scholar]
  18. Fukumaki Y., Shimada K., Takagi Y. Specialized transduction of colicin E1 DNA in Escherichia coli K-12. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3238–3242. doi: 10.1073/pnas.73.9.3238. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gayda R. C., Yamamoto L. T., Markovitz A. Second-site mutations in capR (lon) strains of Escherichia coli K-12 that prevent radiation sensitivity and allow bacteriophage lambda to lysogenize. J Bacteriol. 1976 Sep;127(3):1208–1216. doi: 10.1128/jb.127.3.1208-1216.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Gottesman M. E., Yarmolinsky M. B. Integration-negative mutants of bacteriophage lambda. J Mol Biol. 1968 Feb 14;31(3):487–505. doi: 10.1016/0022-2836(68)90423-3. [DOI] [PubMed] [Google Scholar]
  21. Gottesman S., Zipser D. Deg phenotype of Escherichia coli lon mutants. J Bacteriol. 1978 Feb;133(2):844–851. doi: 10.1128/jb.133.2.844-851.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Guyer M. S., Reed R. R., Steitz J. A., Low K. B. Identification of a sex-factor-affinity site in E. coli as gamma delta. Cold Spring Harb Symp Quant Biol. 1981;45(Pt 1):135–140. doi: 10.1101/sqb.1981.045.01.022. [DOI] [PubMed] [Google Scholar]
  23. Hansen J. B., Abiko Y., Curtiss R., 3rd Characterization of the Streptococcus mutans plasmid pva318 cloned into Escherichia coli. Infect Immun. 1981 Mar;31(3):1034–1043. doi: 10.1128/iai.31.3.1034-1043.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Hobom G., Hogness D. S. The role of recombination in the formation of circular oligomers of the lambda plasmid. J Mol Biol. 1974 Sep 5;88(1):65–87. doi: 10.1016/0022-2836(74)90295-2. [DOI] [PubMed] [Google Scholar]
  25. Hohn B., Collins J. A small cosmid for efficient cloning of large DNA fragments. Gene. 1980 Nov;11(3-4):291–298. doi: 10.1016/0378-1119(80)90069-4. [DOI] [PubMed] [Google Scholar]
  26. Hohn B. DNA as substrate for packaging into bacteriophage lambda, in vitro. J Mol Biol. 1975 Oct 15;98(1):93–106. doi: 10.1016/s0022-2836(75)80103-3. [DOI] [PubMed] [Google Scholar]
  27. Holt R. G., Abiko Y., Saito S., Smorawinska M., Hansen J. B., Curtiss R., 3rd Streptococcus mutans genes that code for extracellular proteins in Escherichia coli K-12. Infect Immun. 1982 Oct;38(1):147–156. doi: 10.1128/iai.38.1.147-156.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Ish-Horowicz D., Burke J. F. Rapid and efficient cosmid cloning. Nucleic Acids Res. 1981 Jul 10;9(13):2989–2998. doi: 10.1093/nar/9.13.2989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Jaskunas S. R., Lindahl L., Nomura M. Identification of two copies of the gene for the elongation factor EF-Tu in E. coli. Nature. 1975 Oct 9;257(5526):458–462. doi: 10.1038/257458a0. [DOI] [PubMed] [Google Scholar]
  30. Kellenberger G., Symonds N., Arber W. Host specificity of DNA produced by Escherichia coli. 8. Its acquisition by phage lambda and its persistence through consecutive growth cycles. Z Vererbungsl. 1966;98(3):247–256. [PubMed] [Google Scholar]
  31. Kleckner N., Barker D. F., Ross D. G., Botstein D. Properties of the translocatable tetracycline-resistance element Tn10 in Escherichia coli and bacteriophage lambda. Genetics. 1978 Nov;90(3):427–461. doi: 10.1093/genetics/90.3.427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. LENNOX E. S. Transduction of linked genetic characters of the host by bacteriophage P1. Virology. 1955 Jul;1(2):190–206. doi: 10.1016/0042-6822(55)90016-7. [DOI] [PubMed] [Google Scholar]
  33. Lindenmaier W., Hauser H., de Wilke I. G., Schütz G. Gene shuttling: moving of cloned DNA into and out of eukaryotic cells. Nucleic Acids Res. 1982 Feb 25;10(4):1243–1256. doi: 10.1093/nar/10.4.1243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Lodish H. F. Secondary structure of bacteriophage f2 ribonucleic acid and the initiation of in vitro protein biosynthesis. J Mol Biol. 1970 Jun 28;50(3):689–702. doi: 10.1016/0022-2836(70)90093-8. [DOI] [PubMed] [Google Scholar]
  35. Maloy S. R., Nunn W. D. Selection for loss of tetracycline resistance by Escherichia coli. J Bacteriol. 1981 Feb;145(2):1110–1111. doi: 10.1128/jb.145.2.1110-1111.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Miwa T., Matsubara K. Formation of oligomeric structures from plasmid DNA carrying cos lambda that is packaged into bacteriophage lambda heads. J Bacteriol. 1983 Jan;153(1):100–108. doi: 10.1128/jb.153.1.100-108.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Mullins J. I., Casey J. W., Nicolson M. O., Burck K. B., Davidson N. Sequence arrangement and biological activity of cloned feline leukemia virus proviruses from a virus-productive human cell line. J Virol. 1981 May;38(2):688–703. doi: 10.1128/jvi.38.2.688-703.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Murchison H., Larrimore S., Curtiss R., 3rd Isolation and characterization of Streptococcus mutans mutants defective in adherence and aggregation. Infect Immun. 1981 Dec;34(3):1044–1055. doi: 10.1128/iai.34.3.1044-1055.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Murray N. E., Brammar W. J., Murray K. Lambdoid phages that simplify the recovery of in vitro recombinants. Mol Gen Genet. 1977 Jan 7;150(1):53–61. doi: 10.1007/BF02425325. [DOI] [PubMed] [Google Scholar]
  40. Sancar A., Hack A. M., Rupp W. D. Simple method for identification of plasmid-coded proteins. J Bacteriol. 1979 Jan;137(1):692–693. doi: 10.1128/jb.137.1.692-693.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Sanderson K. E., Roth J. R. Linkage map of Salmonella typhimurium, Edition VI. Microbiol Rev. 1983 Sep;47(3):410–453. doi: 10.1128/mr.47.3.410-453.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Simon L. D., Gottesman M., Tomczak K., Gottesman S. Hyperdegradation of proteins in Escherichia coli rho mutants. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1623–1627. doi: 10.1073/pnas.76.4.1623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Stahl F. W., McMilin K. D., Stahl M. M., Malone R. E., Nozu Y., Russo V. E. A role for recombination in the production of "free-loader" lambda bacteriophage particles. J Mol Biol. 1972 Jul 14;68(1):57–67. doi: 10.1016/0022-2836(72)90262-8. [DOI] [PubMed] [Google Scholar]
  44. Sternberg N., Tiemeier D., Enquist L. In vitro packaging of a lambda Dam vector containing EcoRI DNA fragments of Escherichia coli and phage P1. Gene. 1977 May;1(3-4):255–280. doi: 10.1016/0378-1119(77)90049-x. [DOI] [PubMed] [Google Scholar]
  45. Umene K., Shimada K., Takagi Y. Packaging of ColE1 DNA having a lambda phage cohesive end site. Mol Gen Genet. 1978 Feb 7;159(1):39–45. doi: 10.1007/BF00401746. [DOI] [PubMed] [Google Scholar]
  46. Uzan M., Danchin A. A rapid test for the rel A mutation in E. coli. Biochem Biophys Res Commun. 1976 Apr 5;69(3):751–758. doi: 10.1016/0006-291x(76)90939-6. [DOI] [PubMed] [Google Scholar]
  47. Vollenweider H. J., Fiandt M., Rosenvold E. C., Szybalski W. Packaging of plasmid DNA containing the cohesive ends of coliphage lambda. Gene. 1980 Apr;9(1-2):171–174. doi: 10.1016/0378-1119(80)90174-2. [DOI] [PubMed] [Google Scholar]
  48. White F. F., Klee H. J., Nester E. W. In vivo packaging of cosmids in transposon-mediated mutagenesis. J Bacteriol. 1983 Feb;153(2):1075–1078. doi: 10.1128/jb.153.2.1075-1078.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Young R. A., Davis R. W. Efficient isolation of genes by using antibody probes. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1194–1198. doi: 10.1073/pnas.80.5.1194. [DOI] [PMC free article] [PubMed] [Google Scholar]

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