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. 1986 Jan;165(1):226–232. doi: 10.1128/jb.165.1.226-232.1986

Genetic and molecular analysis of aroL, the gene for shikimate kinase II in Escherichia coli K-12.

R C DeFeyter, J Pittard
PMCID: PMC214393  PMID: 3001024

Abstract

The gene aroL in Escherichia coli K-12, specifying shikimate kinase II, was contransduced with proC at a frequency of 99%. The gene order is lac proC aroL. A 2.7-kilobase BamHI fragment containing aroL+ was cloned into pBR322. This plasmid conferred highly elevated levels of shikimate kinase synthesis which were subject to repression control by tyrR. The aroL gene was localized within a 730-base-pair region by both subcloning and insertional mutagenesis with Tn1000. A second gene, designated aroM and encoding a protein of molecular weight 26,000, is cotranscribed with aroL. Transcription proceeds in the order aroL aroM in a clockwise direction on the chromosome. The function of aroM remains unknown.

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Selected References

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  1. Bachmann B. J. Linkage map of Escherichia coli K-12, edition 7. Microbiol Rev. 1983 Jun;47(2):180–230. doi: 10.1128/mr.47.2.180-230.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berlyn M. B., Giles N. H. Organization of enzymes in the polyaromatic synthetic pathway: separability in bacteria. J Bacteriol. 1969 Jul;99(1):222–230. doi: 10.1128/jb.99.1.222-230.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  4. Brown K. D., Doy C. H. Transport and utilization of the biosynthetic intermediate shikimic acid in Escherichia coli. Biochim Biophys Acta. 1976 May 28;428(3):550–562. doi: 10.1016/0304-4165(76)90183-5. [DOI] [PubMed] [Google Scholar]
  5. Brown K. D., Somerville R. L. Repression of aromatic amino acid biosynthesis in Escherichia coli K-12. J Bacteriol. 1971 Oct;108(1):386–399. doi: 10.1128/jb.108.1.386-399.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Camakaris H., Pittard J. Autoregulation of the tyrR gene. J Bacteriol. 1982 Apr;150(1):70–75. doi: 10.1128/jb.150.1.70-75.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Camakaris H., Pittard J. Regulation of tyrosine and phenylalanine biosynthesis in Escherichia coli K-12: properties of the tyrR gene product. J Bacteriol. 1973 Sep;115(3):1135–1144. doi: 10.1128/jb.115.3.1135-1144.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Casadaban M. J., Cohen S. N. Lactose genes fused to exogenous promoters in one step using a Mu-lac bacteriophage: in vivo probe for transcriptional control sequences. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4530–4533. doi: 10.1073/pnas.76.9.4530. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Casadaban M. J. Fusion of the Escherichia coli lac genes to the ara promoter: a general technique using bacteriophage Mu-1 insertions. Proc Natl Acad Sci U S A. 1975 Mar;72(3):809–813. doi: 10.1073/pnas.72.3.809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Covarrubias L., Cervantes L., Covarrubias A., Soberón X., Vichido I., Blanco A., Kupersztoch-Portnoy Y. M., Bolivar F. Construction and characterization of new cloning vehicles. V. Mobilization and coding properties of pBR322 and several deletion derivatives including pBR327 and pBR328. Gene. 1981 Jan-Feb;13(1):25–35. doi: 10.1016/0378-1119(81)90040-8. [DOI] [PubMed] [Google Scholar]
  12. DeFeyter R. C., Davidson B. E., Pittard J. Nucleotide sequence of the transcription unit containing the aroL and aroM genes from Escherichia coli K-12. J Bacteriol. 1986 Jan;165(1):233–239. doi: 10.1128/jb.165.1.233-239.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. DeFeyter R. C., Pittard J. Purification and properties of shikimate kinase II from Escherichia coli K-12. J Bacteriol. 1986 Jan;165(1):331–333. doi: 10.1128/jb.165.1.331-333.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Deutch A. H., Smith C. J., Rushlow K. E., Kretschmer P. J. Escherichia coli delta 1-pyrroline-5-carboxylate reductase: gene sequence, protein overproduction and purification. Nucleic Acids Res. 1982 Dec 11;10(23):7701–7714. doi: 10.1093/nar/10.23.7701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ely B., Pittard J. Aromatic amino acid biosynthesis: regulation of shikimate kinase in Escherichia coli K-12. J Bacteriol. 1979 Jun;138(3):933–943. doi: 10.1128/jb.138.3.933-943.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Figurski D. H., Meyer R. J., Helinski D. R. Suppression of Co1E1 replication properties by the Inc P-1 plasmid RK2 in hybrid plasmids constructed in vitro. J Mol Biol. 1979 Sep 25;133(3):295–318. doi: 10.1016/0022-2836(79)90395-4. [DOI] [PubMed] [Google Scholar]
  17. Guyer M. S. The gamma delta sequence of F is an insertion sequence. J Mol Biol. 1978 Dec 15;126(3):347–365. doi: 10.1016/0022-2836(78)90045-1. [DOI] [PubMed] [Google Scholar]
  18. Hershfield V., Boyer H. W., Yanofsky C., Lovett M. A., Helinski D. R. Plasmid ColEl as a molecular vehicle for cloning and amplification of DNA. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3455–3459. doi: 10.1073/pnas.71.9.3455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Im S. W., Davidson H., Pittard J. Phenylalanine and tyrosine biosynthesis in Escherichia coli K-12: mutants derepressed for 3-deoxy-D-arabinoheptulosonic acid 7-phosphate synthetase (phe), 3-deoxy-D-arabinoheptulosonic acid 7-phosphate synthetase (tyr), chorismate mutase T-prephenate dehydrogenase, and transaminase A. J Bacteriol. 1971 Oct;108(1):400–409. doi: 10.1128/jb.108.1.400-409.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Inouye H., Michaelis S., Wright A., Beckwith J. Cloning and restriction mapping of the alkaline phosphatase structural gene (phoA) of Escherichia coli and generation of deletion mutants in vitro. J Bacteriol. 1981 May;146(2):668–675. doi: 10.1128/jb.146.2.668-675.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Klein R. D., Selsing E., Wells R. D. A rapid microscale technique for isolation of recombinant plasmid DNA suitable for restriction enzyme analysis. Plasmid. 1980 Jan;3(1):88–91. doi: 10.1016/s0147-619x(80)90037-2. [DOI] [PubMed] [Google Scholar]
  22. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  23. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  24. Lynn S. P., Cohen L. K., Kaplan S., Gardner J. F. RsaI: a new sequence-specific endonuclease activity from Rhodopseudomonas sphaeroides. J Bacteriol. 1980 May;142(2):380–383. doi: 10.1128/jb.142.2.380-383.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. MONOD J., COHEN-BAZIRE G., COHN M. Sur la biosynthèse de la beta-galactosidase (lactase) chez Escherichia coli; la spécificité de l'induction. Biochim Biophys Acta. 1951 Nov;7(4):585–599. doi: 10.1016/0006-3002(51)90072-8. [DOI] [PubMed] [Google Scholar]
  26. Mattern I. E., Pittard J. Regulation of tyrosine biosynthesis in Escherichia coli K-12: isolation and characterization of operator mutants. J Bacteriol. 1971 Jul;107(1):8–15. doi: 10.1128/jb.107.1.8-15.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Morrison D. A. Transformation in Escherichia coli: cryogenic preservation of competent cells. J Bacteriol. 1977 Oct;132(1):349–351. doi: 10.1128/jb.132.1.349-351.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nakata A., Peterson G. R., Brooks E. L., Rothman F. G. Location and orientation of the phoA locus on the Escherichia coli K-12 linkage map. J Bacteriol. 1971 Sep;107(3):683–689. doi: 10.1128/jb.107.3.683-689.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. PITTARD J. EFFECT OF INTEGRATED SEX FACTOR ON TRANSDUCTION OF CHROMOSOMAL GENES IN ESCHERICHIA COLI. J Bacteriol. 1965 Mar;89:680–686. doi: 10.1128/jb.89.3.680-686.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Pittard J., Wallace B. J. Gene controlling the uptake of shikimic acid by Escherichia coli. J Bacteriol. 1966 Oct;92(4):1070–1075. doi: 10.1128/jb.92.4.1070-1075.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Selker E., Brown K., Yanofsky C. Mitomycin C-induced expression of trpA of Salmonella typhimurium inserted into the plasmid ColE1. J Bacteriol. 1977 Jan;129(1):388–394. doi: 10.1128/jb.129.1.388-394.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. So M., Gill R., Falkow S. The generation of a ColE1-Apr cloning vehicle which allows detection of inserted DNA. Mol Gen Genet. 1975 Dec 30;142(3):239–249. doi: 10.1007/BF00425649. [DOI] [PubMed] [Google Scholar]
  34. Wallace B. J., Pittard J. Regulator gene controlling enzymes concerned in tyrosine biosynthesis in Escherichia coli. J Bacteriol. 1969 Mar;97(3):1234–1241. doi: 10.1128/jb.97.3.1234-1241.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Whipp M. J., Pittard A. J. Regulation of aromatic amino acid transport systems in Escherichia coli K-12. J Bacteriol. 1977 Nov;132(2):453–461. doi: 10.1128/jb.132.2.453-461.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Yagil E., Bracha M., Silberstein N. Further genetic mapping of the phoA-phoR region for alkaline phosphatase synthesis in Escherichia coli K 12. Mol Gen Genet. 1970;109(1):18–26. doi: 10.1007/BF00334043. [DOI] [PubMed] [Google Scholar]
  37. Young I. G., Jaworowski A., Poulis M. I. Amplification of the respiratory NADH dehydrogenase of Escherichia coli by gene cloning. Gene. 1978 Sep;4(1):25–36. doi: 10.1016/0378-1119(78)90012-4. [DOI] [PubMed] [Google Scholar]

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