Fluorescence-based method for selection of recombinant plasmids

A fluorescence-based method for the selection of recombinant plasmids has been developed. Escherichia coli strains bearing plasmids for the overexpression of the gene encoding uroporphyrinogen III methyltransferase (cobA or cysG gene) accumulate fluorescent porphyrinoid compounds. When illuminated w...

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Bibliographic Details
Published inBioTechniques Vol. 19; no. 5; p. 760
Main Authors Roessner, C A, Scott, A I
Format Journal Article
LanguageEnglish
Published England 01.11.1995
Subjects
Aminolevulinic Acid - metabolism
DNA, Recombinant
Escherichia coli - genetics
Fluorescence
Gene Expression
Methyltransferases - genetics
Molecular Sequence Data
Plasmids
Polymerase Chain Reaction
Restriction Mapping
Saccharomyces cerevisiae - genetics
Salmonella typhimurium - genetics
Ultraviolet Rays
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Summary:A fluorescence-based method for the selection of recombinant plasmids has been developed. Escherichia coli strains bearing plasmids for the overexpression of the gene encoding uroporphyrinogen III methyltransferase (cobA or cysG gene) accumulate fluorescent porphyrinoid compounds. When illuminated with ultraviolet light, the cells fluoresce with a bright red color. Replacement or disruption of the gene with other fragments of DNA results in loss of enzymatic activity and nonfluorescent cells. The detection of the recombinant plasmids can be accomplished without special host strains or expensive chromogenic reagents required for the blue-white screening technique.
ISSN:0736-6205