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Fate of genetically modified maize DNA in the oral cavity and rumen of sheep

Published online by Cambridge University Press:  09 March 2007

Paula S. Duggan ,
Philip A. Chambers ,
John Heritage  and
J. Michael Forbes
Paula S. Duggan
Affiliation:
Division of Microbiology, School of Biochemistry and Molecular Biology, University of Leeds, LS2 9JT, UK
Philip A. Chambers
Affiliation:
Division of Microbiology, School of Biochemistry and Molecular Biology, University of Leeds, LS2 9JT, UK
John Heritage
Affiliation:
Division of Microbiology, School of Biochemistry and Molecular Biology, University of Leeds, LS2 9JT, UK
J. Michael Forbes*
Affiliation:
Centre for Animal Sciences, School of Biology, University of Leeds, Leeds LS2 9JT, UK
*
*Corresponding author: Professor J. M. Forbes, fax +44 113 2333144, email j.m.forbes@leeds.ac.uk
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Abstract

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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.

Keywords

Horizontal gene transferDNA survivalInsect-resistant maizeAntibiotic resistanceTransformation efficiency
Type
Research Article
Information
British Journal of Nutrition , Volume 89 , Issue 2 , February 2003 , pp. 159 - 166
Copyright
Copyright © The Nutrition Society 2003

References

Chiter, A, Forbes, JM & Blair, GE (2000) DNA stability in plant tissues: Implications for the possible transfer of genes from genetically-modified food. FEBS Letters 481, 164–168.CrossRefGoogle ScholarPubMed
Davison, J (1999) Genetic exchange between bacteria in the environment. Plasmid 42, 73–91.CrossRefGoogle ScholarPubMed
De Vries, J, Meier, P & Wackernagel, W (2001) The natural transformation of the soil bacteria Pseudomonas stutzeri and Acinetobacter sp. by transgenic plant DNA strictly depends on homologous sequences in the recipient cells. FEMS Letters 195, 211–215.CrossRefGoogle ScholarPubMed
De Vries, J & Wackernagel, W (1998) Detection of npt II (kana-mycin resistance) genes in genomes of transgenic plants by marker rescue transformation. Molecular and General Genetics 257, 606–613.CrossRefGoogle Scholar
Duggan, PS, Chambers, PA, Heritage, J & Forbes, JM (2000) Survival of free DNA encoding antibiotic resistance from transgenic maize and the transformation activity of DNA in silage effluent, ovine saliva and ovine rumen fluid. FEMS Letters 191, 71–77.CrossRefGoogle ScholarPubMed
Faichney, GJ (1993) Digesta Flow. In Quantitative Aspects of Ruminant Digestion and Metabolism, pp. 53–85. [Forbes, JM and France, J, editors]. Wallingford, Oxon: CAB International.Google Scholar
Flint, HJ (1994) Molecular genetics of obligate anaerobes from the rumen. FEMS letters 121, 259–268.CrossRefGoogle ScholarPubMed
Gebhard, F & Smalla, K (1988) Transformation of Acinetobacter sp. strain BD413 by transgenic sugar beet DNA. Applied and Environmental Microbiology 64, 1550–1554.CrossRefGoogle Scholar
Greisen, K, Loeffelholz, M, Purohit, A & Lenog, D (1994) PCR primers for the 16S rRNA gene of most species of pathogenic bacteria, including bacteria found in cerebrospinal fluid. Journal of Clinical Microbiology 32, 335–351.CrossRefGoogle ScholarPubMed
Hanahan, D (1983) Studies on transformation of E. coli with plasmids. Journal of Molecular Biology 166, 557–580.CrossRefGoogle ScholarPubMed
Hupfer, C, Hotzel, H, Sachse, K & Engel, KH (1997) Detection of genetically modified insect-resistant Bt maize by means of polymerase reaction. Zeitschrift für Lebensmittel-Unter-suchung und Forschung 205, 442–445.CrossRefGoogle Scholar
Hupfer, C, Hotzel, H, Sachse, K & Engel, KH (1998) Detection of the genetic modification in heat-treated products of Bt maize by polymerase chain reaction. Zeitschrift für Lebensmittel-Untersuchung und Forschung 206, 203–207.CrossRefGoogle Scholar
Hupfer, C, Mayer, J, Hotzel, H, Sachse, K & Engel, KH (1999) The effect of ensiling on PCR-based detection of genetically modified Bt maize. European Food Research and Technology 209, 301–304.CrossRefGoogle Scholar
Koziel, MG, Beland, GL, Bowman, C, Carozzi, NB, Crenshaw, R, Crossland, L, Dawson, J, Desai, N, Hill, M, Kadwell, S, Launis, K, Maddox, D, Mcpherson, K, Meghji, MR, Merlin, E, Rhodes, R, Warren, GW, Wright, M & Evola, SV (1993) Field performance of elite transgenic maize plants expressing an insecticidal protein derived from Bacillus thuringiensis. Bio-Technology 11, 194–200.Google Scholar
Kuramitsu, HK & Trapa, V (1984) Genetic exchange between oral Streptococci during mixed growth. Journal of General Microbiology 130, 2497–2500.Google ScholarPubMed
Li, YH, Lau, PCY, Lee, JH, Ellen, RP & Cvitkovitch, DG (2001) Natural genetic transformation of Streptococcus mutans growing in biofilms. Journal of Bacteriology 183, 897–908.CrossRefGoogle ScholarPubMed
Lorenz, G & Wackernagel, W (1994) Bacterial gene transfer by natural genetic transformation in the environment. Microbiology Reviews 58, 563–602.CrossRefGoogle ScholarPubMed
Lunsford, RD (1998) Streptococcal transformation: essential features and applications of a natural gene exchange system. Plasmid 39, 10–20.CrossRefGoogle Scholar
Mabilat, C, Goussard, S, Sougakoff, W, Spencer, RC & Courvalin, P (1990) Direct sequencing of the amplified structural gene and promoter for the extended-broad-spectrum b-lactamase TEM-9 (RHH-1) of Klebsiella pneumoniae. Plasmid 23, 27–34.CrossRefGoogle Scholar
Martìn-Orúe, SM, O'Donnell, AG, Ariño, J, Netherwood, T, Gilbert, HJ & Mathers, JC (2002) Degradation of transgenic DNA from genetically modified soya and maize in human intestinal simulations. British Journal of Nutrition 87, 533–542.CrossRefGoogle ScholarPubMed
Méjean, V & Claverys, JP (1993) DNA processing during entry in transformation of Streptococcus pneumoniae. Journal of Biological Chemistry 268, 5594–5599.CrossRefGoogle ScholarPubMed
Mercer, DK, Scott, KP, Bruce-Johnson, WA, Glover, LA & Flint, HJ (1999 a) Fate of free DNA and transformation of the oral bacterium. Streptococcus gordonii DL1 by plasmid DNA in human saliva. Applied & Environmental Microbiology 65, 6–10.CrossRefGoogle ScholarPubMed
Mercer, DK, Melville, CM, Scott, KP & Flint, HJ (1999 b) Natural transformation of the rumen bacterium Streptococcus bovis JB1. FEMS Microbiology Letters 179, 485–490.CrossRefGoogle ScholarPubMed
Mercer, DK, Scott, KP, Melville, CM, Glover, LA & Flint, HJ (2001) Transformation of an oral bacterium via chromosomal integration of free DNA in the presence of human saliva. FEMS Microbiology Letters 200, 163–167.CrossRefGoogle ScholarPubMed
Morrison, M (1996) Do ruminal bacteria exchange genetic material? Journal of Dairy Science 79, 1476–1486.CrossRefGoogle ScholarPubMed
Nielsen, KM, Bones, AM, Smalla, K & van Elsas, JD (1988) Horizontal gene transfer from transgenic plants to terrestrial bacteria – a rare event? Microbiology Reviews 22, 79–103.Google Scholar
Ruiz, TR, Andrews, S & Smith, GB (2000) Identification and characterisation of nuclease activities in anaerobic environmental samples. Canadian Journal of Microbiology 46, 736–740.CrossRefGoogle ScholarPubMed
Russell, JB & Wilson, DB (1988) Potential opportunities and problems for genetically altered rumen microorganisms. Journal of Nutrition 118, 271–279.CrossRefGoogle ScholarPubMed
Sambrook, J, Fritsch, E & Maniatis, T (1989) In Molecular Cloning: a Laboratory Manual, pp. A.1.80–A.1.81 [Ford, N and Nolan, C, editors]. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.Google Scholar
Schubbert, R, Lettmann, C & Doerfler, W (1994) Ingested foreign (phage M13) DNA survives transiently in the gastrointestinal tract and enters the bloodstream of mice. Molecular and General Genetics 242, 495–504.CrossRefGoogle ScholarPubMed
Schubbert, R, Renz, D, Schmitz, B & Doerfler, W (1997) Foreign (M13) DNA ingested by mice reaches peripheral leukocytes, spleen, and liver via the intestinal wall mucosa and can be covalently linked to mouse DNA. Proceedings of the National Academy of Sciences USA 94, 961–966.CrossRefGoogle ScholarPubMed
Westergren, G & Emilson, CG (1983) Prevalence of transformable Streptoccus mutans in human dental plaque. Infection and Immunity 41, 1386–1388.CrossRefGoogle Scholar