Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978

Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid.

A C Chang, S N Cohen
PMCID: PMC222365  PMID: 149110

Abstract

Construction and characterization of a class of multicopy plasmid cloning vehicles containing the replication system of miniplasmid P15A are described. The constructed plasmids have cleavage sites within antibiotic resistance genes for a variety of commonly employed site-specific endonucleases, permitting convenient use of the insertional inactivation procedure for the selection of clones that contain hybrid DNA molecules. Although the constructed plasmids showed DNA sequence homology with the ColE1 plasmid within the replication region, were amplifiable by chloramphenicol or spectinomycin, required DNA polymerase I for replication, and shared other replication properties with ColE1, they were nevertheless compatible with ColE1. P15A-derived plasmids were not self-transmissible and were mobilized poorly by Hfr strains; however, mobilization was complemented by the presence of a ColE1 plasmid within the same cell.

Full text

PDF
1142

Images in this article

1145Image
on p.1145
1147Image
on p.1147
1149Image
on p.1149
1152Image
on p.1152

Selected References

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

  1. Armstrong K. A., Hershfield V., Helinski D. R. Gene cloning and containment properties of plasmid Col E1 and its derivatives. Science. 1977 Apr 8;196(4286):172–174. doi: 10.1126/science.322277. [DOI] [PubMed] [Google Scholar]
  2. Bachmann B. J. Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev. 1972 Dec;36(4):525–557. doi: 10.1128/br.36.4.525-557.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bazaral M., Helinski D. R. Replication of a bacterial plasmid and an episome in Escherichia coli. Biochemistry. 1970 Jan 20;9(2):399–406. doi: 10.1021/bi00804a029. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Blattner F. R., Williams B. G., Blechl A. E., Denniston-Thompson K., Faber H. E., Furlong L., Grunwald D. J., Kiefer D. O., Moore D. D., Schumm J. W. Charon phages: safer derivatives of bacteriophage lambda for DNA cloning. Science. 1977 Apr 8;196(4286):161–169. doi: 10.1126/science.847462. [DOI] [PubMed] [Google Scholar]
  6. Blin N., von Gabain A., Bujard H. Isolation of large molecular weight DNA from agarose gels for further digestion by restriction enzymes. FEBS Lett. 1975 Apr 15;53(1):84–86. doi: 10.1016/0014-5793(75)80688-0. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Bujard H., Mazaitis A. J., Bautz E. K. The size of the rII region of bacteriophage T4. Virology. 1970 Nov;42(3):717–723. doi: 10.1016/0042-6822(70)90317-x. [DOI] [PubMed] [Google Scholar]
  9. CAVALLI L. L., LEDERBERG J., LEDERBERG E. M. An infective factor controlling sex compatibility in Bacterium coli. J Gen Microbiol. 1953 Feb;8(1):89–103. doi: 10.1099/00221287-8-1-89. [DOI] [PubMed] [Google Scholar]
  10. Cabello F., Timmis K., Cohen S. N. Replication control in a composite plasmid constructed by in vitro linkage of two distinct replicons. Nature. 1976 Jan 29;259(5541):285–290. doi: 10.1038/259285a0. [DOI] [PubMed] [Google Scholar]
  11. Carroll D., Brown D. D. Adjacent repeating units of Xenopus laevis 5S DNA can be heterogeneous in length. Cell. 1976 Apr;7(4):477–486. doi: 10.1016/0092-8674(76)90199-9. [DOI] [PubMed] [Google Scholar]
  12. Chang S., Cohen S. N. In vivo site-specific genetic recombination promoted by the EcoRI restriction endonuclease. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4811–4815. doi: 10.1073/pnas.74.11.4811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Clark A. J., Chamberlin M., Boyce R. P., Howard-Flanders P. Abnormal metabolic response to ultraviolet light of a recombination deficient mutant of Escherichia coli K12. J Mol Biol. 1966 Aug;19(2):442–454. doi: 10.1016/s0022-2836(66)80015-3. [DOI] [PubMed] [Google Scholar]
  14. Clewell D. B., Helinski D. R. Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an opern circular DNA form. Proc Natl Acad Sci U S A. 1969 Apr;62(4):1159–1166. doi: 10.1073/pnas.62.4.1159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Clewell D. B. Nature of Col E 1 plasmid replication in Escherichia coli in the presence of the chloramphenicol. J Bacteriol. 1972 May;110(2):667–676. doi: 10.1128/jb.110.2.667-676.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Cohen S. N., Chang A. C., Boyer H. W., Helling R. B. Construction of biologically functional bacterial plasmids in vitro. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3240–3244. doi: 10.1073/pnas.70.11.3240. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Cohen S. N., Chang A. C., Hsu L. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110–2114. doi: 10.1073/pnas.69.8.2110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Cohen S. N., Chang A. C. Recircularization and autonomous replication of a sheared R-factor DNA segment in Escherichia coli transformants. Proc Natl Acad Sci U S A. 1973 May;70(5):1293–1297. doi: 10.1073/pnas.70.5.1293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Cohen S. N., Chang A. C. Revised interpretation of the origin of the pSC101 plasmid. J Bacteriol. 1977 Nov;132(2):734–737. doi: 10.1128/jb.132.2.734-737.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Cohen S. N., Miller C. A. Non-chromosomal antibiotic resistance in bacteria. II. Molecular nature of R-factors isolated from Proteus mirabilis and Escherichia coli. J Mol Biol. 1970 Jun 28;50(3):671–687. doi: 10.1016/0022-2836(70)90092-6. [DOI] [PubMed] [Google Scholar]
  21. Cozzarelli N. R., Kelly R. B., Kornberg A. A minute circular DNA from Escherichia coli 15. Proc Natl Acad Sci U S A. 1968 Jul;60(3):992–999. doi: 10.1073/pnas.60.3.992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Curtiss R., 3rd, Charamella L. J., Berg C. M., Harris P. E. Kinetic and genetic analyses of D-cycloserine inhibition and resistance in Escherichia coli. J Bacteriol. 1965 Nov;90(5):1238–1250. doi: 10.1128/jb.90.5.1238-1250.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Dougan G., Sherratt D. The transposon Tn1 as a probe for studying ColE1 structure and function. Mol Gen Genet. 1977 Mar 7;151(2):151–160. doi: 10.1007/BF00338689. [DOI] [PubMed] [Google Scholar]
  24. FREDERICQ P., BETZ-BAREAU M. Transfert génétique de la propriété colicinogène chez E. coli. C R Seances Soc Biol Fil. 1953 Jun;147(11-12):1110–1112. [PubMed] [Google Scholar]
  25. Garfin D. E., Goodman H. M. Nucleotide sequences at the cleavage sites of two restriction endonucleases from Hemophilus parainfluenzae. Biochem Biophys Res Commun. 1974 Jul 10;59(1):108–116. doi: 10.1016/s0006-291x(74)80181-6. [DOI] [PubMed] [Google Scholar]
  26. Goebel W. Replication of the DNA of the colicinogenic factor E 1 (Col E 1 ) at the restrictive temperature in a DNA replication mutant thermosensitive for DNA polymerase. 3. Nat New Biol. 1972 May 17;237(72):67–70. doi: 10.1038/newbio237067a0. [DOI] [PubMed] [Google Scholar]
  27. Goebel W., Schrempf H. Isolation of minicircular deoxyribonucleic acids from wild strains of Escherichia coli and their relationship to other bacterial plasmids. J Bacteriol. 1972 Sep;111(3):696–704. doi: 10.1128/jb.111.3.696-704.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Hamer D. H., Thomas C. A., Jr Molecular cloning of DNA fragments produced by restriction endonucleases Sa1I and BamI. Proc Natl Acad Sci U S A. 1976 May;73(5):1537–1541. doi: 10.1073/pnas.73.5.1537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Hedgpeth J., Goodman H. M., Boyer H. W. DNA nucleotide sequence restricted by the RI endonuclease. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3448–3452. doi: 10.1073/pnas.69.11.3448. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Helinski D. R., Herschman H. R. Effect of Rec mutations on the activity of colicinogenic factors. J Bacteriol. 1967 Sep;94(3):700–706. doi: 10.1128/jb.94.3.700-706.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Hershfield V., Boyer H. W., Chow L., Helinski D. R. Characterization of a mini-ColC1 plasmid. J Bacteriol. 1976 Apr;126(1):447–453. doi: 10.1128/jb.126.1.447-453.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Inselburg J. Replication of colicin E1 plasmid DNA in minicells from a unique replication initiation site. Proc Natl Acad Sci U S A. 1974 Jun;71(6):2256–2259. doi: 10.1073/pnas.71.6.2256. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Inselburg J. Studies of colicin E1 plasmid functions by analysis of deletions and TnA insertions of the plasmid. J Bacteriol. 1977 Oct;132(1):332–340. doi: 10.1128/jb.132.1.332-340.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Kopecko D. J., Cohen S. N. Site specific recA--independent recombination between bacterial plasmids: involvement of palindromes at the recombinational loci. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1373–1377. doi: 10.1073/pnas.72.4.1373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Kretschmer F. J., Chang A. C., Cohen S. N. Indirect selection of bacterial plasmids lacking identifiable phenotypic properties. J Bacteriol. 1975 Oct;124(1):225–231. doi: 10.1128/jb.124.1.225-231.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Kretschmer P. J., Cohen S. N. Selected translocation of plasmid genes: frequency and regional specificity of translocation of the Tn3 element. J Bacteriol. 1977 May;130(2):888–899. doi: 10.1128/jb.130.2.888-899.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Landy A., Ruedisueli E., Robinson L., Foeller C., Ross W. Digestion of deoxyribonucleic acids from bacteriophage T7, lambda, and phi 80h with site-specific nucleases from Hemophilus influenzae strain Rc and strain Rd. Biochemistry. 1974 May 7;13(10):2134–2142. doi: 10.1021/bi00707a022. [DOI] [PubMed] [Google Scholar]
  38. Leder P., Tiemeier D., Enquist L. EK2 derivatives of bacteriophage lambda useful in the cloning of DNA from higher organisms: the lambdagtWES system. Science. 1977 Apr 8;196(4286):175–177. doi: 10.1126/science.322278. [DOI] [PubMed] [Google Scholar]
  39. Lobban P. E., Kaiser A. D. Enzymatic end-to end joining of DNA molecules. J Mol Biol. 1973 Aug 15;78(3):453–471. doi: 10.1016/0022-2836(73)90468-3. [DOI] [PubMed] [Google Scholar]
  40. Lovett M. A., Katz L., Helinski D. R. Unidirectional replication of plasmid ColE1 DNA. Nature. 1974 Sep 27;251(5473):337–340. doi: 10.1038/251337a0. [DOI] [PubMed] [Google Scholar]
  41. Messing J., Staudenbauer W. L., Hofschneider P. H. Inhibition of minicircular DNA replication in Escherichia coli 15 by rifampicin. Nat New Biol. 1972 Aug 16;238(85):202–203. doi: 10.1038/newbio238202a0. [DOI] [PubMed] [Google Scholar]
  42. Monk M., Kinross J. 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):971–978. doi: 10.1128/jb.109.3.971-978.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Murray N. E., Murray K. Manipulation of restriction targets in phage lambda to form receptor chromosomes for DNA fragments. Nature. 1974 Oct 11;251(5475):476–481. doi: 10.1038/251476a0. [DOI] [PubMed] [Google Scholar]
  44. Oka A., Takanami M. Cleavage map of colicin E1 plasmid. Nature. 1976 Nov 11;264(5582):193–196. doi: 10.1038/264193a0. [DOI] [PubMed] [Google Scholar]
  45. Polisky B., Greene P., Garfin D. E., McCarthy B. J., Goodman H. M., Boyer H. W. Specificity of substrate recognition by the EcoRI restriction endonuclease. Proc Natl Acad Sci U S A. 1975 Sep;72(9):3310–3314. doi: 10.1073/pnas.72.9.3310. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Rambach A., Tiollais P. Bacteriophage lambda having EcoRI endonuclease sites only in the nonessential region of the genome. Proc Natl Acad Sci U S A. 1974 Oct;71(10):3927–3930. doi: 10.1073/pnas.71.10.3927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Sharp P. A., Cohen S. N., Davidson N. Electron microscope heteroduplex studies of sequence relations among plasmids of Escherichia coli. II. Structure of drug resistance (R) factors and F factors. J Mol Biol. 1973 Apr 5;75(2):235–255. doi: 10.1016/0022-2836(73)90018-1. [DOI] [PubMed] [Google Scholar]
  48. Sharp P. A., Sugden B., Sambrook J. Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose--ethidium bromide electrophoresis. Biochemistry. 1973 Jul 31;12(16):3055–3063. doi: 10.1021/bi00740a018. [DOI] [PubMed] [Google Scholar]
  49. Tait R. C., Rodriguez R. L., Boyer H. W. Altered tetracycline resistance in pSC101 recombinant plasmids. Mol Gen Genet. 1977 Mar 16;151(3):327–331. doi: 10.1007/BF00268797. [DOI] [PubMed] [Google Scholar]
  50. Thomas M., Cameron J. R., Davis R. W. Viable molecular hybrids of bacteriophage lambda and eukaryotic DNA. Proc Natl Acad Sci U S A. 1974 Nov;71(11):4579–4583. doi: 10.1073/pnas.71.11.4579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Timmis K., Cabello F., Cohen S. N. Cloning, isolation, and characterization of replication regions of complex plasmid genomes. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2242–2246. doi: 10.1073/pnas.72.6.2242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Timmis K., Cabello F., Cohen S. N. Utilization of two distinct modes of replication by a hybrid plasmid constructed in vitro from separate replicons. Proc Natl Acad Sci U S A. 1974 Nov;71(11):4556–4560. doi: 10.1073/pnas.71.11.4556. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Tomizawa J., Sakakibara Y., Kakefuda T. Replication of colicin E1 plasmid DNA in cell extracts. Origin and direction of replication. Proc Natl Acad Sci U S A. 1974 Jun;71(6):2260–2264. doi: 10.1073/pnas.71.6.2260. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Ullrich A., Shine J., Chirgwin J., Pictet R., Tischer E., Rutter W. J., Goodman H. M. Rat insulin genes: construction of plasmids containing the coding sequences. Science. 1977 Jun 17;196(4296):1313–1319. doi: 10.1126/science.325648. [DOI] [PubMed] [Google Scholar]
  55. Warren G., Sherratt D. Complementation of transfer deficient ColE1 mutants. Mol Gen Genet. 1977 Mar 7;151(2):197–201. doi: 10.1007/BF00338695. [DOI] [PubMed] [Google Scholar]
  56. von Gabain A., Hayward G. S., Bujard H. Physical mapping of the HindIII, EcoRI, Sal and Sma restriction endonuclease cleavage fragments from bacteriophage T5 DNA. Mol Gen Genet. 1976 Feb 2;143(3):279–290. doi: 10.1007/BF00269404. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES