On the fate of plant or other foreign genes upon the uptake in food or after intramuscular injection in mice
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Uptake and persistence of the DNA of bacteriophage M13 and the cloned gene for the green fluorescent protein (GFP) as test genes for food-ingested DNA have previously been traced from the intestinal contents, via the gut wall, Peyer's patches and peripheral white blood cells to spleen and liver, and via the placenta to fetuses and newborn animals. We have now chosen a natural scenario and fed soybean leaves to mice. The distribution of the plant-specific, nucleus-encoded ribulose-1,5-bisphosphate carboxylase (Rubisco) gene has been studied in the mouse. The Rubisco gene or fragments of it can be recovered in the intestine from 2 h up to 49 h after feeding, and in the cecum up to 121 h after ingestion. Thus, plant-associated, naturally fed DNA is more stable in the intestinal tract than naked DNA. Rubisco gene-specific PCR products have also been amplified from spleen and liver DNA. There is no evidence for the expression of orally administered genes, as assessed by the RT-PCR method. Moreover, mice have been continuously fed daily with GFP DNA for 8 generations and have been examined for the transgenic state by assaying DNA isolated from tail tips, occasionally from internal organs of the animals, by PCR. The results have been uniformly negative and argue against the germline transfer of orally administered DNA. Upon the intramuscular injection of GFP DNA, authentic GFP DNA fragments have been amplified by PCR from DNA from muscle for up to 17 months post-injection, and from DNA from organs remote from the site of injection up to 24 h post injection. GFP fragments can also be retrieved from the intestinal contents up to 6 h post injection. The organism apparently eliminates injected foreign DNA via the liver-bile-intestinal route.
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