抗草甘膦大豆转基因pcr检测及其饲用安全研究-动物营养与饲料科学专业毕业论文.docxVIP

AbstractThis Abstract This paper was conducted to study PCR methods for detecting exotic genes from genetically modified(GM)RR glyphosate-tolerant soybeans，soybearl meal and feedstuffs，and to make a safety assessment for its application in animals． Qualitative and quantitative PCR methods for detecting GM soybeans and its derivative feedstuflg were developed．Firstly genomic DNA was obtained wi也CTAB method or kit for extraction from GMO．After qualitative PCR was used to screening and ampli母different target genes in the exotic gene cassette，the PCR products were recovered and cut fnrther with the corresponding restrictive enzyme． The limit of detection was 0．1％with qualitative PCR．When nested PCR was employed．as little as Pg of soybcan template DNA could be detected．Based on the detection wj也double—primer PCR．the soybearls and feedstaffs from America and Argentina were GM positive．In addition，real—time quantitative PCR method was developed witIl lluorescence dye SYBR Green 1 what combined to doublerstrand DNA．The standard curves of／ec and CaMV35s g：e】／les were drawn with O．9997 and 0．9992 ofR2 value respectively．The relative deviation was within 5-11％between the determinated and actual values ofknown levels ofstandard or simulated soybeans． A series of experiments including growth of weanling piglets，digestion of grower pigs and subchronic toxity of rats were conducted to evaluate feeding value and safety ofsoybean meal from GM RR glyphosate-tolerant soybcans．The analyzed proximate nutrient，amino acids，trace mineral，fatty acid and anti-nutrition factors such as trypsin inhibitor in RK so#earls or soybean meal were within the normal range．RR soybearl meal had no significant effects on performance and blood physiological and biochemical parameters of weanling piglets．The experiment of grower fired with a T—carmula at the distal ileum showed that ileal apparent and true digestibility of crude protein and amino acids were not difierent between the conventional and RR soyb