Fate of genetically modified maize DNA in the oral cavity and rumen of sheep.
The polymerase chain reaction (PCR) technique was used to investigate the fate of a transgene in the rumen of sheep fed silage and maize grains from an insect-resistant maize line. A 1914-bp DNA fragment containing the entire coding region of the synthetic cryIA(b) gene was still amplifiable from rumen fluid sampled 5 h after feeding maize grains. The same target sequence, however, could not be amplified from rumen fluid sampled from sheep fed silage prepared from the genetically modified maize line. PCR amplification of a shorter (211-bp), yet still highly specific, target sequence was possible with rumen fluid sampled up to 3 and 24 h after feeding silage and maize grains, respectively. These findings indicate that intact transgenes from silage are unlikely to survive significantly in the rumen since a DNA sequence 211-bp long is very unlikely to transmit genetic information. By contrast, DNA in maize grains persists for a significant time and may, therefore, provide a source of transforming DNA in the rumen. In addition, we have examined the biological activity of plasmid DNA that had previously been exposed to the ovine oral cavity. Plasmid extracted from saliva sampled after incubation for 8 min was still capable of transforming competent Escherichia coli to kanamycin resistance, implying that DNA released from the diet within the mouth may retain sufficient biological activity for the transformation of competent oral bacteria.
Duggan
Paula S
PS
Division of Microbiology, School of Biochemistry and Molecular Biology, University of Leeds, LS2 9JT, UK.
Chambers
Philip A
PA
Heritage
John
J
Michael Forbes
J
J
D000821:Animal Feed; D000818:Animals; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics*; D001427:Bacterial Toxins; D032461:Chromosomes, Plant; D004247:DNA / Q000032:analysis; D004731:Endotoxins / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D022761:Gene Transfer, Horizontal; D006460:Hemolysin Proteins; D007613:Kanamycin Resistance / Q000235:genetics; D009055:Mouth / Q000382:microbiology; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction / Q000379:methods; D012417:Rumen / Q000382:microbiology; D012756:Sheep / Q000378:metabolism*; D013997:Time Factors; D014169:Transformation, Bacterial / Q000235:genetics*; D019076:Transgenes; D003313:Zea mays / Q000235:genetics*
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004731:Endotoxins; D006460:Hemolysin Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis; D004247:DNA