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. 1987 Apr;169(4):1777–1779. doi: 10.1128/jb.169.4.1777-1779.1987

IS2 activates the ilvA gene of Pseudomonas cepacia in Escherichia coli.

G Barsomian, T G Lessie
PMCID: PMC212018  PMID: 3031023

Abstract

The ilvA gene of Pseudomonas cepacia was expressed poorly in Escherichia coli. Insertion of IS2 upstream of the cloned gene dramatically increased its transcription, resulting in an 85-fold increase in threonine dehydratase (deaminase) specific activity. The results confirm earlier reports that IS2 promotes efficient expression of foreign genes in E. coli.

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Selected References

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  1. Beckman W., Gaffney T., Lessie T. G. Correlation between auxotrophy and plasmid alteration in mutant strains of Pseudomonas cepacia. J Bacteriol. 1982 Mar;149(3):1154–1158. doi: 10.1128/jb.149.3.1154-1158.1982. [DOI] [PMC free article] [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. Brachet P., Eisen H., Rambach A. Mutations of coliphage lambda affecting the expression of replicative functions O and P. Mol Gen Genet. 1970;108(3):266–276. doi: 10.1007/BF00283357. [DOI] [PubMed] [Google Scholar]
  4. Brosius J., Walz A. DNA sequences flanking an E. coli insertion element IS2 in a cloned yeast TRP5 gene. Gene. 1982 Feb;17(2):223–228. doi: 10.1016/0378-1119(82)90076-2. [DOI] [PubMed] [Google Scholar]
  5. Carmichael G. G., McMaster G. K. The analysis of nucleic acids in gels using glyoxal and acridine orange. Methods Enzymol. 1980;65(1):380–391. doi: 10.1016/s0076-6879(80)65049-6. [DOI] [PubMed] [Google Scholar]
  6. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dmochowska A., Berse B., Kowalska I., Skrzypek M., Wegleński P. Expression of the Aspergillus nidulans argB gene in Escherichia coli. J Gen Microbiol. 1986 Jun;132(6):1729–1737. doi: 10.1099/00221287-132-6-1729. [DOI] [PubMed] [Google Scholar]
  8. Ghosal D., Sommer H., Saedler H. Nucleotide sequence of the transposable DNA-element IS2. Nucleic Acids Res. 1979 Mar;6(3):1111–1122. doi: 10.1093/nar/6.3.1111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Lessie T. G., Whiteley H. R. Properties of threonine deaminase from a bacterium able to use threonine as sole source of carbon. J Bacteriol. 1969 Nov;100(2):878–889. doi: 10.1128/jb.100.2.878-889.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. McCorkle G. M., Leathers T. D., Umbarger H. E. Physical organization of the ilvEDAC genes of Escherichia coli strain K-12. Proc Natl Acad Sci U S A. 1978 Jan;75(1):89–93. doi: 10.1073/pnas.75.1.89. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Meyers J. A., Sanchez D., Elwell L. P., Falkow S. Simple agarose gel electrophoretic method for the identification and characterization of plasmid deoxyribonucleic acid. J Bacteriol. 1976 Sep;127(3):1529–1537. doi: 10.1128/jb.127.3.1529-1537.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Southern E. Gel electrophoresis of restriction fragments. Methods Enzymol. 1979;68:152–176. doi: 10.1016/0076-6879(79)68011-4. [DOI] [PubMed] [Google Scholar]
  13. Thomas P. S. Hybridization of denatured RNA transferred or dotted nitrocellulose paper. Methods Enzymol. 1983;100:255–266. doi: 10.1016/0076-6879(83)00060-9. [DOI] [PubMed] [Google Scholar]
  14. Walz A., Ratzkin B., Carbon J. Control of expression of a cloned yeast (Saccharomyces cerevisiae) gene (trp5) by a bacterial insertion element (IS2). Proc Natl Acad Sci U S A. 1978 Dec;75(12):6172–6176. doi: 10.1073/pnas.75.12.6172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Wong H. C., Allenza P., Lessie T. G. Hydroxyamino acid utilization and alpha-ketobutyrate toxicity in Pseudomonas cepacia. J Bacteriol. 1980 Oct;144(1):441–443. doi: 10.1128/jb.144.1.441-443.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Wood A. G., Konisky J. Activation of expression of a cloned archaebacterial gene in Escherichia coli by IS2, IS5, or deletions. Mol Gen Genet. 1985;198(2):309–314. doi: 10.1007/BF00383012. [DOI] [PubMed] [Google Scholar]

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