[1]刘冰,张英,张海波,等.大豆中转基因成分筛查策略研究[J].大豆科学,2016,35(03):411-417.[doi:10.11861/j.issn.1000-9841.2016.03.0411]
LIU Bing,ZHANG Ying,ZHANG Hai-bo,et al.Screening Strategy of Genetically Modified Soybean Based on QualitativePCR Methods[J].Soybean Science,2016,35(03):411-417.[doi:10.11861/j.issn.1000-9841.2016.03.0411]
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《大豆科学》[ISSN:1000-9841/CN:23-1227/S]
卷:
第35卷
期数:
2016年03期
页码:
411-417
栏目:
出版日期:
2016-05-20
- Title:
-
Screening Strategy of Genetically Modified Soybean Based on QualitativePCR Methods
- 作者:
-
刘冰; 张英; 张海波; 杨娟妮; 陈西; 张田
-
陕西省种子管理站/农业部农作物种子质量监督检验测试中心( 西安) ,陕西西安710018
- Author(s):
-
LIU Bing; ZHANG Ying; ZHANG Hai-bo; YANG Juan-ni; CHEN Xi; ZHANG Tian
-
( Shaanxi Seed Administration Bureau / The Crop Seed Quality Testing Center,Xi’an 710018,China)
-
- 关键词:
-
转基因大豆; 筛查检测; PCR 方法; 灵敏度
- Keywords:
-
Genetically modified soybean; Screening method; PCR method; Sensitivity
- 分类号:
-
S565. 1
- DOI:
-
10.11861/j.issn.1000-9841.2016.03.0411
- 文献标志码:
-
A
- 摘要:
-
转基因大豆是目前我国进口量最大的转基因作物,为避免转基因大豆及制品的违法使用,快速筛查大豆中转基因成分的方法策略亟待开发。为检索已知商业化转基因大豆的外源基因信息,通过构建转基因大豆常用转化元件的数据库,建立了转基因大豆的筛查策略。结果表明: 已知的15 个转基因大豆转化事件中,利用CaMV35S 启动子、CP4-epsps 基因、Bt 基因、pat 基因4 个靶标元件的组合,可筛查12 个转基因大豆的转化事件; 另有3 个转化事件需要使用转化事件特异性引物检测。使用4 个靶标元件和转化事件组合的筛查方法检测的灵敏度可达到1 g·kg - 1 ,可对大豆中的转化事件进行快速、高灵敏度的筛查。
- Abstract:
-
Genetically modified ( GM) soybean is the largest amount crop of imported genetically modified organism in China.In order to avoid unauthorized genetically modified ( GM) soybean events occurred occasionally,a quick screening method isneeded urgently. According to the inserted gene elements information of commercial domestic and foreign in GM soybean events,a screening strategy was developed for detecting complex molecular structure in GM soybean events based on elementfrequencies. A combination of CaMv35S promoter,CP4-epsps /Bt /Bar /pat genes as screening target elements could screentwelve genetically modified ( GM) soybean events,the event specific primer could detect other three genetically modified( GM) soybean. Results showed that the detection sensitivity of this method was 1 g·kg - 1. In conclusion,the screening strategyis currently a unique and very valuable tool which could quickly screen out GM soybean detection with high sensitivity andspecificity.
参考文献/References:
[1] 侯文胜,林抗雪,陈普,等. 大豆规模化转基因技术体系的构建及其应用[J]. 中国农业科学,2014,47 ( 21 ) : 4198-4210.( Hou W S,Lin K X,Chen P,et al. Establishment and prospectof efficient transformation systems for soybean [J]. Scientia AgriculturaSinica,2014,47( 21) : 4198-4210.
[2] James C. 2014 年全球生物技术/转基因作物商业化发展态势[J]. 中国生物工程杂志,2015,35 ( 1 ) : 1-14. ( James C.Global status of commercialized biotech /GM crops: 2014 [J].China Biotechnology,2015,35( 1) : 1-14. )
[3] 殷瑞锋,徐雪高. 2014 年大豆市场形势分析与2015 年展望[J]. 农业展望,2014 ( 12) : 4-13. Yin R F,Xu X G. Soybeanmarket in 2014 and its prospect for 2015[J]. Agriculture Outlook,2014 ( 12) : 4-13.
[4] GM Crop Database of Center for Environmental Risk Assessment,http: / /cera-gmc. org /GM. CropDatabase. [2015-07-30].
[5] GMO Database of GMO Compass,EU,http: / /www. gmo-compass.org /eng /gmo /db. [2015-07-30].
[6] http: / /gmdd. shgmo. org. GMO Detection Method Database( GMDD) ,http: / /gmdd. shgmo. org. [2015-07-30].
[7] 中华人民共和国农业部. 进口用作加工原料的农业转基因生物审批情况[OL]. http: / /www. moa. gov. cn /ztzl /zjyqwgz /spxx /Ministry of Agriculture of the People’s Republic of China. The approvalof imported genetically modified plants used for processingmaterials. http: / /www. moa. gov. cn /ztzl /zjyqwgz /spxx /.
[8] 中华人民共和国海关总署. 2014 年12 月全国进口重点商品量值表[OL]. http: / /www. customs. gov. cn /publish /portal0 /tab49666 /info729731. htm General Administration of Customs ofthe People’s Republic ofChina. The national key value of goodsimported in December 2014. http: / /www. customs. gov. cn /publish/portal0 /tab49666 /info729731. htm.
[9] 孟素芬. 中国进口转基因大豆使用管理研究[D]. 郑州: 河南工业大学,2012. ( Meng S F. The using management research ofimported transgenic soybeans of China[D]. Zhengzhou: Henan Universityof Technology,2012. )
[10] Zel J,Cankar K,Ravnikar M,et al. Accreditation of GMO detectionlaboratories: Improving the reliability of GMO detection[J].Accreditation and Quality Assurance,2006,10( 10) : 531-536.
[11] 张丽,武玉花,吴刚,等. 转基因油菜筛查检测策略研究[J].中国油料作物学报,2012,34 ( 1) : 74-81. ( Zhang L,Wu Y H,Wu G,et al. Strategy of transgenic rapeseed screening based onexogenous gene elements[J]. Chinese Journal of Oil Crop Sciences,2012,34( 1) : 74-81.
[12] 余婧,张孝廉,赵丹,等. 烟草转基因研究常用遗传元件筛查策略[J]. 中国烟草学报,2014,20( 1) : 78-83. ( Yu J,ZhangX L,Zhao D, et al. Screening strategy of genetic elements used intransgenic study[J]. Acta Tabacaria Sinica,2014,20 ( 1) : 78-83. )
[13] 农业部. 农业部1782 号公告-3-2012 转基因植物及其产品成分检测调控元件CaMV 35S 启动子、FMV 35S 启动子、NOS 启动子、NOS 终止子和CaMV 35S 终止子定性PCR 方法[S]. 北京: 农业部,2012. ( Ministry of Agriculture. Announcement by theMinistry of Agriculture,No. 1782-3-2012. Detection of geneticallymodified plants and derived products-Qualitative PCR methods forthe regulatory elements CaM V 35S promoter,FM V 35S promoter,NOS promoter,NOS terminator and CaM V 35S terminator[S].Beijing: Ministry of Agriculture,2012. )
[14] 农业部. 农业部953 号公告-6-2007 转基因植物及其产品成分检测抗虫转Bt 基因水稻定性PCR 方法[S]. 北京: 农业部,2007. Ministry of Agriculture. Announcement by the Ministry ofAgriculture,No. 953-6-2007. Detection of genetically modifiedplants and their derived products -Qualitative PCR methods for Btrice to control insect pests[S]. Beijing: Ministry of Agriculture,2007. )
[15] 农业部. 农业部1782 号公告-6-2012 转基因植物及其产品成分检测bar 或pat 基因定性PCR 方法[S]. 北京: 农业部,2012. ( Ministry of Agriculture. Announcement by the Ministry ofAgriculture,No. 1782-6-2012. Detection of genetically modifiedplants and derived products-Qualitative PCR method of bar or patgene[S]. Beijing: Ministry of Agriculture,2012. )
[16] 农业部. 农业部1861 号公告-5-2012 转基因植物及其产品成分检测CP4-epsps 基因定性PCR 方法[S]. 北京: 农业部,2012. ( Ministry of Agriculture. Announcement by the Ministry ofAgriculture,No. 1861-5-2012. Detection of genetically modifiedplants and derived products Qualitative PCR method for CP4-epspsgene[S]. Beijing: Ministry of Agriculture,2012. )
[17] 农业部. 农业部1782 号公告-4-2012 转基因植物及其产品成分检测高油酸大豆305423 及其衍生品种定性PCR 方法[S].北京: 农业部,2012. ( Ministry of Agriculture. Announcement bythe Ministry of Agriculture,No. 1782-4-2012. Detection of geneticallymodified plants and derived products-Qualitative PCR methodfor high oleic acid soybean 305423 and its derivates[S]. Beijing:Ministry of Agriculture,2012. )
[18] 农业部. 农业部1782 号公告-1-2012 转基因植物及其产品成分检测耐除草剂大豆356043 及其衍生品种定性PCR 方法[S ] . 北京: 农业部,2012. ( Ministry of Agriculture.Announcement by the Ministry of Agriculture,No. 1782-1-2012. Detection of genetically modified plants and derived products-Qualitative PCR method for for herbicide-tolerant soybean356043 and its derivates[S]. Beijing: Ministry of Agriculture,2012. )
[19] 农业部. 农业部1782 号公告-5-2012 转基因植物及其产品成分检测耐除草剂大豆CV127 及其衍生品种定性PCR 方法[S]. 北京: 农业部, 2012. ( Ministry of Agriculture.Announcement by the Ministry of Agriculture,No. 1782-5-2012. Detection of genetically modified plants and derived products-Qualitative PCR method for herbicide-tolerant soybean CV127and its derivates[S]. Beijing: Ministry of Agriculture,2012. )
[20] 吴明生,云晓敏,宋歌,等. 转基因玉米种子快速筛查方法研究与应用[J]. 生物技术通报,2013 ( 1) : 102-106. ( Wu M S,Yun X M,Song G,et al. Study and application of a rapid screeningmethod for GM maize seeds[J]. Biotechnology Bulletin,2013( 1) : 102-106. )
[21] Wu Y H,Wang Y L,Li J,et al. Development of a general methodfor detection and quantification of the P35S promoter based onassessment of existing methods[J]. Scientific Reports,2014( 4) :7358. )
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ZHANG Bin-bin,LI Yong-guang,GAI Jiang-nan,et al.Distance and Frequency of Gene Flow in Transgenic Soybean Overexpressing TaDREB3[J].Soybean Science,2011,30(03):563.[doi:10.11861/j.issn.1000-9841.2011.04.0563]
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