The Detection Of The Transgenic Soybean GTS40-3-2 Related Genes And Their Products Of Livestock Products Sold And Toxicology Study Of Sprague-Dawley Rats (Rattus Norvegicus)

[Objective] Genetically modified (GM) crops are planted widely to promote the global food production, alleviate the problem to the environment due to the use of pesticides, reduce poverty and hunger, bring ongoing considerable interest for the global environment (especially in the developing countries and regions), economic and social benefits. However, biological security problems from GM crops, as of the date of birth is accompanied by, the debate has never stopped. Although GM crops and their products must go through a series of strict toxicology before commercial certification, the current GM safety test is limited to animal testing, and most of the animal feeds only take short-term observation. Feeding on the long-term impact of chronic lack of experimental results and data, lack of effective argument for the influence of human body. In this paper, to provide data support for our GM feed forage safety evaluation system and have important theoretical and realistic significance from the reasonable application of transgenic livestock products by feeding animals producing or GM sources of raw materials products in China, the GM animal feed composition were detected, the people's Daily diet of meat, eggs, milk and oil were researched and chronic SD rats were feeded for a long time.[Method] The experiment used the qualitative PCR technology to detect the 30 laying layers and pigs feed on the market of GM ingredients in Shanxi, overlapping PCR method (using the gene cloning, transformation and expression) to build the standard plasmid, standard plasmid molecules constructed to detect GM ingredients of agriculture products such as meat, eggs, milk, oil and protein by qualitative and quantitative PCR and ELISA in the Shanxi market, tissue biopsies, hematology and blood biochemistry indexes, quantitative and qualitative PCR and ELISA to explore the toxic effects and residue of the transgenic soybean GTS40-3-2 on SD rats by long-term feeding animal model.[Result] (1) 30 feeds were checked in the Shanxi market (including 20 different feed of layers and pigs in different developmental stages from different manufacturer. Feed varieties were representative and wide coverage. There were 25 feed samples test results were positive (positive share of 83.3%). But there was only one marked "genetically modified" ingredients. GM product identification rate was only 4%. The results showed that the layer and pig feeds mostly contained soybean transformation event GTS40-3-2, a small amount of feeds contained maize transformant event with MON810.(2) Four genes had been successfully cloned into a plasmid carrier molecules by using the method of molecular cloning (qualitative PCR and overlapping PCR technology), which was used as the positive standard in subsequent qualitative and quantitative PCR detection.(3) Meat, eggs, milk and oil related gene and protein of transgenic biological product constituents were detected in Shanxi market. All animal feeds from 11 farmers contained GM ingredients, especially containing soybean specific transformant event GTS40-3-2. Then experiment collected the animal faces from 11 farmers and kept on detection related gene, all faces were only checked out 18SrDNA. Ten broiler were detected themetabolic residues of transgenic soybean GTS40-3-2 from a farmhouse on the spot check by qualitative PCR, quantitative PCR and protein detection. Caecum contents and intestinal fecal of broilers were checked out GM ingredients, other solid organs werenot found containing GM ingredients. In addition, edible oil, pork, eggs and milk were carried on the related detection of protein and genes in the Shanxi market. The intestinal contents of pig were only found to contain genetically modified ingredients, other solid organ were not detected GMO related genes and proteins.14 cooking oil detected out 13 oil samples containing different content of soybean internal Lectin gene, only two oil samples were detect out very low GTS40-3-2 gene copies.(4) The experiment detected the body gain, feed consumption, hematology indexes, blood biochemical indicators and pathological observation indicators of the first generation and the second generation SD rat by adding containing soybean transformant event GTS40-3-2 pulp, but it was not found that GM and non-GM groups had significant difference statistically significant. By qualitative PCR, quantitative PCR and ELISA test, the experiment detected the genes of intestinal flora and viscera associated with the feed, only the rat intestinal dung and cecum contents were detected residues of GM ingredients, the rat intestinal flora and solid organ had been found residue of GM ingredients.[Conclusion] By qualitative PCR, real-time PCR, slicing technique and protein ELISA detection method, animal feeds, pig organs, broiler viscera, eggs, milk, cooking oil and long-term feeding SD rats were tested in the Shanxi market. The results showed that the animal feeds contained universally GM ingredients. But livestock products such as pork, broiler, eggs and milk related feed had not been found GM ingredients and protein residues by qualitative PCR, quantitative PCR and ELISA detection,. Cooking oil test results showed there were low copies of both internal and external source genes.