Construction of green fluorescent protein-based vectors for genetic analysis in bacteria

Authors

  • A. SUÁREZ
  • A. GÜTTLER
  • M. STRÄTZ
  • L.H. STAENDNER
  • K.M. TIMMIS
  • C.A. GUZMÁN

Keywords:

Vectors., Promoter-probing, Minitransposon Tn5, Oreen fluorescent protein.

Abstract

The green fluorescent protein (GFP) gene, gfp, was used to develop versatile reporter systems for genetic analysis in, and monitoring of bacteria. The reporter system is available on a plasmid and on a mini-transposon located in a suicide delivery plasmid for generation of chromosomal fusions. To achieve sensitivity levels necessary for use in monocopy and for detection of single cells, the 3'-end of gfp was replaced by that of a modified gfp gene characterized by a 4S-fold stronger signal than that exhibited by

the natural OFP, and the modified gfp gene equipped with the strong translation signals of the atpE gene. Transfer of the minitrasnposon into two different Pseudomonas spp. and Alcaligenes eutrophus produced random chromosomal fusions, some 5% of which exhibited fluorescence detectable by eye. Individual OFP+ cells were readily observed by fluorescence microscopy. The detailed map of the step-by-step construction is depicted.

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References

CODY, C. W., PRASHER, D. C., WESTLER, W. M., PRENDERGAST, F. G. Y WARD, W. W.: "Chemical structure of the hexapeptide chromophore of the Aequorea green-fluorescent protein". B iochemistry (I993), 32: 1212-1218.
CHALFIE, M., TU, Y., EUSKIRCHEN, G., WARD, W. W. Y PRASHER, D. c.: "Green fluorescent protein as a marker for gene expression". Science (1994) 263 :802-805 .

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Published

1997-03-20

How to Cite

1.
SUÁREZ A, GÜTTLER A, STRÄTZ M, STAENDNER L, TIMMIS K, GUZMÁN C. Construction of green fluorescent protein-based vectors for genetic analysis in bacteria . Ars Pharm [Internet]. 1997 Mar. 20 [cited 2024 May 19];38(2-3):319-28. Available from: https://revistaseug.ugr.es/index.php/ars/article/view/21219

Issue

Vol. 38 No. 2-3 (1997)

Section

Original Articles

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