| Since the commercial planting of genetically modified crops in 1996,the rapid development of the global genetically modified crops industry.The safe of the genetically modified food is payed attention,in order to meet the consumer’s right to know,the right to choose and strengthen the safety management of the study and application of GMOs,countries have introduced the corresponding GMO safety management laws and regμlations to modify crops and products.The establishment of an accurate detection method is the prerequisite for the safety management of genetically modified organisms,and the reference material is an indispensable material base in the detection technology.The transgenic plasmid standard as one of the standard substance,the industry was a genetically modified gold standard material ".Transgenic Papaya Lines Event 55-1 and "Huanong No.1" have been planted began commercial cultivation in 1996 and 2006.But China has not been a corresponding testing standard,and standard material is lackly.This study is based on two kinds of transgenic papaya Event 55-1 and "Huanong No.1" as the research objects,try to construct standard molecule for detection of transgenic papaya positive plasmid,and detect its application in the detection of genetically modified plants,as positive standard substance of transgenic detection to provide reference for development and application.The main results obtained in this study are as follows:1.Through the search of public databases and literature search,through evaluation,to find and confirm the nucleotide sequence used to construct plasmid.2.Successfully constructing internal standard contains papaya genes papain,Chy,specific sequence of transgenic papaya "Event 55-1",transgenic papaya "Huanong No.1" specific sequence of positive plasmid molecμle.3.Analyze the sensitivity and specificity of analysis and alternative method of positive plasmid molecules,analysis results showed that the positive plasmid has good specificity,can replace the matrix material as a positive control,qualitative PCR detection limit of1000 copies.4.Used flesh papaya,papaya leaves,papaya seeds,dry papaya and preserved fruit as detection object,test positive plasmid molecules application in detection of transgenic papaya,and field planting of transgenic papaya,a preliminary survey of sales of transgenic papaya products in the market.The results showed that the positive plasmid molecules could be used as the positive control. |
