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1、PCRsZEyjT_吴影1, 2,陆徐忠1,赵伟1, 汪秀峰1, 李莉1null(1.8jS,g230031; 2.jv3S,g230036)K1null 以转基因大豆a水稻等样品为材料,采用多重PCR技术对转基因作物中常见的启动子a终止子a筛选标记基因和转入的目的基因等多个外源转基因元件进行检测,以期建立一套快速a准确a高效的转基因农作物筛查鉴定技术b通过多重扩增实验,建立了关于转基因大豆检测的大豆内源Lectin基因,花椰菜花叶病毒(Cauliflower mosaic virus,CaMV)35S启动子和根癌农杆菌胭脂碱合成酶基因NOS终止子的三重PCR分析体系,及关于转基因水稻检测的C
2、aMV35SaNPTII和HPT 基因的三重PCR分析的技术体系b并以大豆a水稻等农作物样品为检测材料,采用单一PCR和多重PCR技术同时进行检测,结果表明多重PCR方法具有快速a高效a简便a准确等特点,在转基因成分的检测上具有非常重要的实际应用价值和潜力b1oMnull 转基因成分;多重PCR;检测;技术体系ms|null Q943.2null nullDSMnull Anull nullcI|null 0517- 6611(2006)07- 1297- 03Analysis Methodof GeneticallyModifiedDetectionof Multiplex Polymera
3、se ChainReaction(MPCR)WU Ying etal null (Rice Research Institute, AnhuiAcademy of Agricultural Sciences, Hefei, Anhui 230031)Abstractnull The genetically modified ingredients introduced including promoter, terminator, selectable marker genes andstructuralgenes(encodnulling the novel protein) etc wer
4、e detectedwith the multiplex PCR technology in order to establish a quick, exact and effective technical system ofthe screening of genetically modified crops. The triplex PCR of soya inner LECTIN gene, Cauliflower mosaic virus (CaMV)35Spromoter andAnullgronullbacterium tum ef aciens nopaline synthas
5、e (NOS) terminator, and the triplex PCR of CaMV35S promoter, NPTII and HPT genes were sucnullcessfully experimented. The transgenic soya andrice sampleswere tested withthe multiplex and simplex PCR technique, respectively. The renullsults showed the multiplex PCRwas quick, effective, simple and exac
6、t, andmight play an important role inthe detection of genetically modifiedcomponent.Key wordsnull Genetically modified ingredient; Multiplex PCR; Detection; Technical systemnull nullyT9F,m1,2,NH,yT3In3,4,SaxaSttE1pyTLS&,jysX,S=_f_b1pyB*eaVLLZE,LjTySbPCR/Ty_/XWVb-S=X3yAU,eyS:yyy,yNtys9_AH_bYPCR_ysH,T
7、N,rq,O;yBPCRQraL#y_/ZE,Ly_TA1pb6,y,TK1JTB,yT*20W90M7S,7O9y|V?LC,yN7Zyy_EsilBTb3yva,raeLPCR/yT0a0a4S:yyy_,YV9L,yv=LectinaCaMV35S0NOS0yPCRs/8#CaMV35SaNPTIIHPTyPCRs/8bjT.y/8y,rjTnull 国家农业科技成果转化资金项目(2004EA710042)bTenull 吴影(1981- ),女,安徽蚌埠人,硕士研究生,研究方向:植物基因工程bnull nullnull 该研究是在安徽省农科院院长杨剑波研究员的悉心指导下完成的,同时还得
8、到了易成新a张海涛a倪大虎a张毅a皮桃花等老师和同事的支持和帮助blnull 2006null01null163Nahnf4ByaaeLZEb1nullZE1. 1null#nullyvvFluka,yv|vLii,dyv|vgb1. 2nullk4#Nnull Taq DNAadNTP1Promega; 100 bp ladder DNA Marker1Takara;k4SsBbMJResearch PTCnull200PCRN; DY501N;Pharmacia GeneQuant IIN; Bionullrad Gel Docd;Eppendof 5810RTb1. 3nullnulln
9、V1b1. 4null9DNA4|null9DNA!CTABE5,DNAAN(Pharmacia, GeneQuant II|)cBbH|5nulll DNAA,0.8%j_DNA4|b1.5 nullBPCR9null PCRQ8: 1 null PCR buffer, 2. 0mmol MgCl2,0.2mmol dNTPs 2nulll, 0. 5nullmol,DNA100ng, 2U Taq DNA,ddH2O835nulllbPCRQHq:94 null 5min;94 null 30s,|30s,72 null 30s,35;72 null 7minb|NPTII56 nullH
10、PT60. 5 null,(62 nullbPCR2. 0%j_,Gel DocdTsb1.6 nullPCR_8nullPCRQ8: 0.4mmol dNTPs, 3UTaq DNA,saPCRQHq#_ZEnull 1.5nullbnull nullisv=LectinyaCaMV35S0aCP4nullEPSPSyCaMV35SaNPTIIaHPTy2F,i1!4F:1null1null1,2null2null1. 5,1null1.5null2,2null1null1. 5bjS, Journal of Anhui Agri. Sci. 2006, 34(7): 1297- 1299null null null null null null null null null null null null null null null null null3I null 孙红忠null3nnull 孙红忠null nullV1#9vl9y()vlbpLectin LECTIN Forward 5null GCCCTCTACTCCACCCCCATCC 3null 118v=yLECTIN Reverse 5null GCCCATCTGCAAGCCTTTTTGTG
