Potential for gene transfer from recombinant Escherichia coli K-12 used in bovine somatotropin production to indigenous bacteria in river water.
This study examined the transfer of the plasmid pBGH1, an expression vector for bovine somatotropin (BST), from Escherichia coli K-12 strain W3110G [pBGH1] to indigenous microorganisms present in flasks containing Missouri River water. Strain LBB269 is a nalidixic acid-resistant derivative of W3110G which was used as a plasmid-free control strain in these studies. Water samples were inoculated with strains W3110G [pBGH1] and LBB269; after 21 days of incubation the number of viable colony-forming units (CFU) of W3110G [pBGH1] and LBB269 were reduced from an initial level of about 1 x 10(7) CFU per ml to less than 1 CFU per 100 ml. At this time indigenous microbes resistant to both ampicillin and tetracycline (the antibiotic resistance markers on pBGH1) were isolated from 100 ml of water from each of the flasks inoculated with either strain W3110G [pBGH1] or LBB269. Plasmid DNA was isolated from these organisms and examined for sequences containing the gene for BST from pBGH1, using a polymerase chain reaction (PCR) assay. As expected, the day 0 sample from the flask inoculated with E. coli K-12 strain W3110G [pBGH1] gave a positive PCR response and the day 0 sample from the flask inoculated with E. coli K-12 strain LBB269 gave a negative PCR response. All of the day 21 samples containing indigenous microbes isolated from flasks that were inoculated with either W3110G [pBGH1] or LBB269 were negative in the PCR assay, indicating that the target sequence from pBGH1 was not present in any of these indigenous microorganisms.(ABSTRACT TRUNCATED AT 250 WORDS)
Bogosian
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Animal Sciences Division, Monsanto Company, Chesterfield, MO 63198.
Morris
P J
PJ
Weber
D B
DB
Kane
J F
JF
D001483:Base Sequence; D015169:Colony Count, Microbial; D004269:DNA, Bacterial / Q000235:genetics*; D004274:DNA, Recombinant / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development; D005618:Fresh Water; D013006:Growth Hormone; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D013678:Technology, Pharmaceutical; D014162:Transfection; D014871:Water Microbiology
D004269:DNA, Bacterial; D004274:DNA, Recombinant; D013006:Growth Hormone