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 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.
Roessner
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CA
Chemistry Department, Texas A&M University, College Station 77843-3255, USA.
Scott
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AI
D000622:Aminolevulinic Acid / Q000378:metabolism; D004274:DNA, Recombinant; D004926:Escherichia coli / Q000235:genetics; D005453:Fluorescence; D015870:Gene Expression; D008780:Methyltransferases / Q000235:genetics; D008969:Molecular Sequence Data; D010957:Plasmids; D016133:Polymerase Chain Reaction; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics; D012486:Salmonella typhimurium / Q000235:genetics; D014466:Ultraviolet Rays
D004274:DNA, Recombinant; D000622:Aminolevulinic Acid; D008780:Methyltransferases; C060508:uroporphyrin-III C-methyltransferase