Skip to main content
The EMBO Journal logoLink to The EMBO Journal
. 1987 Sep;6(9):2519–2523. doi: 10.1002/j.1460-2075.1987.tb02538.x

Characterization of the herbicide-resistance gene bar from Streptomyces hygroscopicus

Charles J Thompson 1,4, N Rao Movva 1, Richard Tizard 2, Reto Crameri 1,5, Julian E Davies 1,4, Marc Lauwereys 3, Johan Botterman 3
PMCID: PMC553668  PMID: 16453790

Abstract

A gene which confers resistance to the herbicide bialaphos (bar) has been characterized. The bar gene was originally cloned from Streptomyces hygroscopicus, an organism which produces the tripeptide bialaphos as a secondary metabolite. Bialaphos contains phosphinothricin, an analogue of glutamate which is an inhibitor of glutamine synthetase. The bar gene product was purified and shown to be a modifying enzyme which acetylates phosphinothricin or demethylphosphinothricin but not bialaphos or glutamate. The bar gene was subcloned and its nucleotide sequence was determined. Interspecific transfer of this Streptomyces gene into Escherichia coli showed that it could be used as a selectable marker in other bacteria. In the accompanying paper, bar has been used to engineer herbicide-resistant plants.

Keywords: bialaphos, phospinothricin, acetyltransferase, herbicide resistance, bar

Full text

PDF
2519

Selected References

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

  1. Ambartsumyan N. S., Mazo A. M. Elimination of the secondary structure effect in gell sequencing of nucleic acids. FEBS Lett. 1980 Jun 2;114(2):265–268. doi: 10.1016/0014-5793(80)81130-6. [DOI] [PubMed] [Google Scholar]
  2. Atal C. K., Zutshi U., Rao P. G. Scientific evidence on the role of Ayurvedic herbals on bioavailability of drugs. J Ethnopharmacol. 1981 Sep;4(2):229–232. doi: 10.1016/0378-8741(81)90037-4. [DOI] [PubMed] [Google Scholar]
  3. Bayer E., Gugel K. H., Hägele K., Hagenmaier H., Jessipow S., König W. A., Zähner H. Stoffwechselprodukte von Mikroorganismen. 98. Phosphinothricin und Phosphinothricyl-Alanyl-Alanin. Helv Chim Acta. 1972 Jan 31;55(1):224–239. doi: 10.1002/hlca.19720550126. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Block M. D., Botterman J., Vandewiele M., Dockx J., Thoen C., Gosselé V., Movva N. R., Thompson C., Montagu M. V., Leemans J. Engineering herbicide resistance in plants by expression of a detoxifying enzyme. EMBO J. 1987 Sep;6(9):2513–2518. doi: 10.1002/j.1460-2075.1987.tb02537.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Campbell J. L., Richardson C. C., Studier F. W. Genetic recombination and complementation between bacteriophage T7 and cloned fragments of T7 DNA. Proc Natl Acad Sci U S A. 1978 May;75(5):2276–2280. doi: 10.1073/pnas.75.5.2276. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chater K. F., Bruton C. J. Resistance, regulatory and production genes for the antibiotic methylenomycin are clustered. EMBO J. 1985 Jul;4(7):1893–1897. doi: 10.1002/j.1460-2075.1985.tb03866.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Herrera-Estrella L., Block M. D., Messens E., Hernalsteens J. P., Montagu M. V., Schell J. Chimeric genes as dominant selectable markers in plant cells. EMBO J. 1983;2(6):987–995. doi: 10.1002/j.1460-2075.1983.tb01532.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Imai S., Seto H., Sasaki T., Tsuruoka T., Ogawa H., Satoh A., Inouye S., Niida T., Otake N. Studies on the biosynthesis of bialaphos (SF-1293). 4. Production of phosphonic acid derivatives, 2-hydroxyethylphosphonic acid, hydroxymethylphosphonic acid and phosphonoformic acid by blocked mutants of Streptomyces hygroscopicus SF-1293 and their roles in the biosynthesis of bialaphos. J Antibiot (Tokyo) 1984 Nov;37(11):1505–1508. doi: 10.7164/antibiotics.37.1505. [DOI] [PubMed] [Google Scholar]
  11. Imai S., Seto H., Sasaki T., Tsuruoka T., Ogawa H., Satoh A., Inouye S., Niida T., Otake N. Studies on the biosynthesis of bialaphos (SF-1293). 6. Production of N-acetyl-demethylphosphinothricin and N-acetylbialaphos by blocked mutants of Streptomyces hygroscopicus SF-1293 and their roles in the biosynthesis of bialaphos. J Antibiot (Tokyo) 1985 May;38(5):687–690. doi: 10.7164/antibiotics.38.687. [DOI] [PubMed] [Google Scholar]
  12. Kieser T. Factors affecting the isolation of CCC DNA from Streptomyces lividans and Escherichia coli. Plasmid. 1984 Jul;12(1):19–36. doi: 10.1016/0147-619x(84)90063-5. [DOI] [PubMed] [Google Scholar]
  13. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Norrander J., Kempe T., Messing J. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene. 1983 Dec;26(1):101–106. doi: 10.1016/0378-1119(83)90040-9. [DOI] [PubMed] [Google Scholar]
  16. Seto H., Imai S., Tsuruoka T., Ogawa H., Satoh A., Sasaki T., Otake N. Studies on the biosynthesis of bialaphos (SF-1293) Part 3. Production of phosphinic acid derivatives, MP-103, MP-104 and MP-105, by a blocked mutant of Streptomyces hygroscopicus SF-1293 and their roles in the biosynthesis of bialaphos. Biochem Biophys Res Commun. 1983 Mar 29;111(3):1008–1014. doi: 10.1016/0006-291x(83)91400-6. [DOI] [PubMed] [Google Scholar]
  17. Seto H., Imai S., Tsuruoka T., Satoh A., Kojima M., Inouye S., Sasaki T., Otake N. Studies on the biosynthesis of bialaphos (SF-1293). 1. Incorporation of 13C- and 2H-labeled precursors into bialaphos. J Antibiot (Tokyo) 1982 Dec;35(12):1719–1721. doi: 10.7164/antibiotics.35.1719. [DOI] [PubMed] [Google Scholar]
  18. Seto H., Sasaki T., Imai S., Tsuruoka T., Ogawa H., Satoh A., Inouye S., Niida T., Otake N. Studies on the biosynthesis of bialaphos (SF-1293). 2. Isolation of the first natural products with a C-P-H bond and their involvement in the C-P-C bond formation. J Antibiot (Tokyo) 1983 Jan;36(1):96–98. doi: 10.7164/antibiotics.36.96. [DOI] [PubMed] [Google Scholar]
  19. Shah D. M., Horsch R. B., Klee H. J., Kishore G. M., Winter J. A., Tumer N. E., Hironaka C. M., Sanders P. R., Gasser C. S., Aykent S., Siegel N. R., Rogers S. G., Fraley R. T. Engineering herbicide tolerance in transgenic plants. Science. 1986 Jul 25;233(4762):478–481. doi: 10.1126/science.233.4762.478. [DOI] [PubMed] [Google Scholar]
  20. Shaw W. V. Chloramphenicol acetyltransferase from chloramphenicol-resistant bacteria. Methods Enzymol. 1975;43:737–755. doi: 10.1016/0076-6879(75)43141-x. [DOI] [PubMed] [Google Scholar]
  21. Thompson C. J., Gray G. S. Nucleotide sequence of a streptomycete aminoglycoside phosphotransferase gene and its relationship to phosphotransferases encoded by resistance plasmids. Proc Natl Acad Sci U S A. 1983 Sep;80(17):5190–5194. doi: 10.1073/pnas.80.17.5190. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Thompson J., Rae S., Cundliffe E. Coupled transcription--translation in extracts of Streptomyces lividans. Mol Gen Genet. 1984;195(1-2):39–43. doi: 10.1007/BF00332721. [DOI] [PubMed] [Google Scholar]
  23. Vandekerckhove J., Bauw G., Puype M., Van Damme J., Van Montagu M. Protein-blotting on Polybrene-coated glass-fiber sheets. A basis for acid hydrolysis and gas-phase sequencing of picomole quantities of protein previously separated on sodium dodecyl sulfate/polyacrylamide gel. Eur J Biochem. 1985 Oct 1;152(1):9–19. doi: 10.1111/j.1432-1033.1985.tb09157.x. [DOI] [PubMed] [Google Scholar]
  24. Vara J. A., Portela A., Ortín J., Jiménez A. Expression in mammalian cells of a gene from Streptomyces alboniger conferring puromycin resistance. Nucleic Acids Res. 1986 Jun 11;14(11):4617–4624. doi: 10.1093/nar/14.11.4617. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES