Construction Of Reference Molecular For Quantitative PCR Detection And The Effect Of Transgenic Soybean GTS40-3-2 On Liver And Kidney Function In Sprague-Dawley Rats

At present,a variety of transgenic crops has became raw materials,they were used for processing foods and animal feeds.We lack of the standard material of genetically modified detection.The standard of quantitative detection for deep processing of products in our country is not perfect.Most of safety tests about animal feed were limited to the short-term observation.This study aims to provide technical support of molecular building,establish the standard method about quantitative detection of genetically modified components in processed products and feed safety monitoring.By using the bioinformatics methods,we found the sequence of endogenous gene and foreign sources genes in maize.The methods of molecular cloning were used to construct the Standard reference plasmid molecules which can detect genetically modified corn components;For the existed plasmid of soybean and constructed plasmid of corn,we gave them dilute steps,and repeated quantitative PCR reaction three times;We bought some processed products(raw materials were soybeans and corn)on the market.Using four methods,we extracted their DNA;Using plasmid molecular,we drawn standard curves and quantitative detected processed products;Simulating animal model of transgenic soybean GTS40-3-2 long-term feeding on rats,we done some researches which were liver and kidney function index determination,bacteria counting,biopsy and quantitative PCR detected the toxicity effect of genetically modified ingredients the residue on liver and kidney in the third generation male SD rats.Standard reference molecules we successful constructed were recombine plasmid molecules containing nine event-specific sequences and the endogenous gene.The two plasmid molecules had better stability,indicated that they could be successfully used as standard material for quantitative PCR.By comparing different methods,obtained better DNA template;nine of 16 samples had very lower transformant contents,a sample contained three kinds of transformant contents;The experiment detected the organ/body weight ratios,bacteria number,blood biochemistry,liver and kidney histopathology of the third generation male rats which were fed transgenic and non-transgenic soybean,.but it was not found two groups had significant difference statistically.By quantitative detecting,we can not found genetically modified ingredients residues in liver and kidney.Two Standard reference molecules we successful constructed could be used as standard material for detection GM products;Raw materials for processing food were complicated,different products extracted DNA method were different;Preliminary determined genetically modified soybean GTS40-3-2 on SD rats was safety.