Effect Of Three Genetic Engineering Products On Animal Safty

1) For finding especial endogenous gene of transgenic rice,several rices single copy genes selected as the primary candidates for rice endogenous reference gene were obtained from database and library online.Sequences of primary candidate genes were blasted in the database online to analyze their homology with those of other crops. Finally the sequence of sucrose phosphate synthase (SPS) gene with the characters of species specificity and homogeneity in species was selected as the formal candidate endogenous reference gene. Meanwhile, the specific routinely primers, TaqMan primers and probe were designed based on the candidate endogenous reference gene sequence, and employed in conventional PCR and real-time PCR to validate their species specificity and homogeneity in species. In addition, southern blotting method was employed to validate the copy number of SPS gene in different rice varieties. All the results indicated that the candidate gene, SPS was suit for using as one endogenous reference gene in genetically modified rice PCR analysis and the exogenous gene horizontal transformation research during biosafety estimation.2) A rapid and reliable TaqMan quantitative PCR methods to estimate the number of integrated copies was developed, which used rice SPS gene as the endogenous reference gene. Firstly, twenty-six transgenic rice samples which have exogenous GUS and HPT genes both from seventy-two tested rice samples by using qualitative PCR and GUS staining methods. Then specific primers and probes for the exogenous GUS and HPT genes were designed and corresponding PCR cycling conditions suitable for the qualitative and quantitative PCR assays were set up. The detection sensitivities of quantitative PCR were selected, HPT and GUS genes were ten copies. These results demonstrated that this method was suitable for use in the qualitative and quantitative detection of transgenic rice. On the basis of these experiments, the standard curves of endogenous SPS gene and exogenous HPT, GUS genes were generated respectively. Then the copy number of transgenes were estimated using relative quantitative methods by calculating the ratio of endogenous gene copies and exogenous gene copies of the same DNA amounts. These experiments results showed that, among the twenty-six tested samples, there are sixteen and nine samples haveone HPT and GUS gene copy respetively; three and nine samples have two HPT and GUS copies respectively; both three samples have three HPT and GUS copies respectively; four and five samples have four or more HPT and GUS copies respectively. Furthermore, by comparing the copy number of GUS gene and HPT gene, thirty-eight percents of the tested rice samples have occurred rearrangements.3) Thirty-two rats weighing about 150 g were randomly divided into two equal groups. One group was fed with the feedstuff that contained 30 percent transgenic codA rice, another control group was fed with the feedstuff that contained 30 percent non-transgenic rice, there are same nutrient in two feedstuff's. After 30 days feeding, the ratio of rats growth, the level of rats physiological metabolism such as the level of GH, Insulin, Glucagon, T4, T3, TSH etc, and the genetic horizontal transformed were analyzed.The results indicated that the weight of all treated rats were not naught compare with control rats (P>0.05), the level of GH, Insulin, Glucagon, T4, T3 and TSH of treated rats did not have a significant difference with the control, and the SUN, Glucose, GPT and the total cholesterol of treated rats were all same with the control group(P>0.05), no target DNA sequence of transgenic rice endogenous SPS gene or exogenous codA gene were detected in rats different organs except for the gastrointestinal contents, and no genetic horizontal transformed was happened in rats after 30 days feeding.4)To analyze the influence of physiological metabolism and genetic horizontal transformation of rats fed with genetically modified soybean containing CP4EPSPS, 32 rats, each with about 150 g weight, were randomly divided into two groups. One g...