食用糖浆及其原料转基因成分的荧光PCR检测
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摘要
采用加大取样量(取10管共50mL样品)、PBS缓冲液预处理、深加工食品DNA试剂盒核酸提取、富集DNA溶液总体积、加大反应体系DNA模板量的方法,成功提取到了糖浆制品的内源基因,建立了食用糖浆中转基因成分实时荧光PCR定性检测方法.同时,对糖浆原料(玉米淀粉)进行转基因成分检测,结果83.3%样品均检出外源基因成分.
The endogenous gene of edible syrup was extracted successfully in this experiment by increasing sample amount(10 tubes of 50 mL samples),PBS buffer pretreatment,using DNA extraction kit for highly processing of food,enriching the total volume of DNA solution,and increasing the amount of DNA template in the reaction system.Real-time fluorescence quantitative PCR method for the detection of transgenes in edible syrup was established.Genetically modified ingredient of corn starch samples(processed materials of edible syrup)were detected in the process of experiment.The results show that,foreign genes are detected in 83.3% samples.
The endogenous gene of edible syrup was extracted successfully in this experiment by increasing sample amount(10 tubes of 50 mL samples),PBS buffer pretreatment,using DNA extraction kit for highly processing of food,enriching the total volume of DNA solution,and increasing the amount of DNA template in the reaction system.Real-time fluorescence quantitative PCR method for the detection of transgenes in edible syrup was established.Genetically modified ingredient of corn starch samples(processed materials of edible syrup)were detected in the process of experiment.The results show that,foreign genes are detected in 83.3% samples.
引文
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[20]梁克红.饲料加工工艺对大豆转基因成分影响规律的研究[D].北京:中国农业科学院饲料研究所,2009.LIANG K H.Degradation of transgenic ingredients of genetically modified soybean during feed processing[D].Beijing:Chinese Academy of Agricultural Sciences Institute of Feed,2009.
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[22]朱占齐,王卫国.转基因作物安全性及外源基因加工降解研究进展[J].饲料工业,2010,31(10):54-57.ZHU Z Q,WANG W G.Research advancement of gm crop safety and degradation of foreign gene[J].Feed Industry,2010,31(10):54-57.
[23]敖金霞.主要粮食作物深加工制品转基因检测技术研究[D].哈尔滨:东北农业大学,2008.AO J X.Study on transgenic detecting techniques for highly processed products of main crops[D].Harbin:Northeast Agricultural University,2008.
[24]柳方方,董美,李凯,等.转基因成分低水平混杂问题的现状[J].生物技术通报,2017,33(3):1-5.LIU F F,DONG M,LI K,et al.Global status on low level presence of genetically modified crops[J].Biotechnology Bulletin,2017,33(3):1-5.
[2]LERAT S,ENGLAND L S,VINCENT M L,et al.Realtime polymerase chain reaction quantification of the transgenes for roundup ready corn and roundup ready soybean in soil samples[J].Journal of Agricultural&Food Chemistry,2005,53(5):1337-1342.
[3]林清,彭于发,吴红,等.转基因作物及产品检测技术研究进展[J].西南农业学报,2009,22(2):513-517.LIN Q,PENG Y F,WU H,et al.Research progress of detection techniques for genetically modified crops and products[J].Southwest China Journal of Agricultural Sciences,2009,22(2):513-517.
[4]国家认证认可监督管理委员会.蜂蜜中转基因成分检测方法普通PCR方法和实时荧光PCR方法:SN/T 2135—2008[S].北京:中国标准出版社,2008.China Certification&Accreditation Institute.Detection of genetically modified components in honey:contentional PCR and real-time fluorescence PCR:SN/T 2135-2008[S].Beijing:Standards Press of China,2008.
[5]张秀杰,刁青云,宛煜嵩,等.蜂蜜中转基因成分的PCR检测[J].基因组学与应用生物学,2010,9(3):582-587.ZHANG X J,DIAO Q Y,WAN Y S,et al.PCR detection method for genetically modified ingredient in honey[J].Genomics and Applied Biology,2010,9(3):582-587.
[6]贾晨曦,郭晓华,刘广纯.不同保存方式下金龟甲虫DNA提取方法及28SrDNA序列分析[J].沈阳大学学报(自然科学版),2013,25(6):436-440.JIA C X,GUO X H,LIU G C.DNA extraction and28srDNA sequences analysis of scarab beetle specimens preserved under different conditions[J].Journal of Shenyang University(Natural Science),2013,25(6):436-440.
[7]张卓,胡建楠,王晓昕,等.基于线粒体COⅠ~COⅡ序列的黑带食蚜蝇(Episyrphus balteatus)遗传多样性初探[J].沈阳大学学报(自然科学版),2015,27(2):116-119.ZHANG Z,HU J N,WANG X X,et al.Genetic diversity of Episyrphus balteatus based on mitochondrial DNA sequence COⅠ~COⅡ[J].Journal of Shenyang University(Natural Science),2015,27(2):116-119.
[8]程红梅,彭于发,金芫军,等.一种快速、简便提取大豆油DNA的方法及转基因大豆油的检测[J].中国农业科学,2007,40(5):1069-1072.CHENG H M,PENG Y F,JIN Y J,et al.A simple and rapid method for isolation of DNA from detection of transgene sequences in soybean oil[J].Scientia Agriculture Sinica,2007,40(5):1069-1072.
[9]国家认证认可监督管理委员会.植物及其加工产品中转基因成分实时荧光PCR定性检测方法:SN/T 1204-2016[S].北京:中国标准出版社,2017.China Certification&Accreditation Institute.Protocol of the real-time PCR for detecting genetically modified plants and their derived products:SN/T 1204-2016[S].Beijing:Standards Press of China,2017.
[10]邵碧英,王德峰,傅碧忠,等.酱油、烤鳗酱油DNA提取及原料成分的荧光PCR检测[J].食品科学,2009,30(22):240-243.SHAO B Y,WANG D F,FU B Z,et al.Extraction of DNA from soy sauce and roast eel sauce and detection of raw material ingredients using fluorescence PCR[J].Food Science,2009,30(22):240-243.
[11]CHAPELA M J,SOTELO C G,PEREZ-MARTIN R I,et al.Comparison of DNA extraction methods from muscle of canned tuna for species identification[J].Food Control,2007,18(10):1211-1215.
[12]MAFRA I,SILVA S A,MOREIRA E J,et al.Comparative study of DNA extraction methods for soybean derived food products[J].Food Control,2008,19(12):1183-1190.
[13]孙敏,粟志平,梁成珠,等.粉丝类产品中转基因成分的检测[J].食品科学,2008,29(6):230-233.SUN M,SU Z P,LIANG C Z,et al.Detection of genetically modified ingredient in vermicelli[J].Food Science,2008,29(6):230-233.
[14]粟志平,耿金培,王洪来,等.粉丝中植物源性成分的PCR定性检测方法研究[J].检验检疫科学,2004,14(5):7-11.SU Z P,GENG J P,WANG H L,et al.Research of PCR for detecting plant ingredient in vermicelli[J].Inspection and Quarantine Science,2004,14(5):7-11.
[15]索娜,刘佳,刘梅,等.米饭和米酒中Bt基因快速检测方法的建立[J].食品科学,2013,34(12):217-222.SUO N,LIU J,LIU M,et al.Rapid detection of Bt gene in cooked rice and rice wine[J].Food Science,2013,34(12):217-222.
[16]白卫滨,孙建霞,程国灵,等.实时荧光PCR技术定量检测转基因豆粉的研究[J].食品科学,2009,30(6):238-242.BAI W B,SUN J X,CHENG G L,et al.Study on quantitative detection of glyphosate-resistant genetically modified soybean powder by real-time fluorescence PCR method[J].Food Science,2009,30(6):238-242.
[17]JAMES D,SCHMIDT A M,WALL E,et al.Reliable detection and identification of genetically modified maize,soybean,and canola by multiplex PCR analysis.[J].Journal of Agricultural&Food Chemistry,2003,51(20):5829.
[18]金红,赵昕,王永,等.转基因作物深加工产品的DNA提取方法研究[J].华北农学报,2006,21(2):53-56.JIN H,ZHAO X,WANG Y,et al.A DNA isolation method for the qualitative detection of transgenic deeplyprocessed products[J].ACTA Agriculturae BorealiSinica,2006,21(2):53-56.
[19]王爱迪,冉晓华,陈磊,等.超强磁性微球提取深加工转基因食品DNA[J].食品科学,2013,34(10):122-125.WANG A D,RAN X H,CHEN L,et al.A DNA isolation method for deeply processed transgenic foods based on high-magnetic response microspheres[J].Food Science,2013,34(10):122-125.
[20]梁克红.饲料加工工艺对大豆转基因成分影响规律的研究[D].北京:中国农业科学院饲料研究所,2009.LIANG K H.Degradation of transgenic ingredients of genetically modified soybean during feed processing[D].Beijing:Chinese Academy of Agricultural Sciences Institute of Feed,2009.
[21]张兴敏,于洪敏,魏健,等.转基因食品中外源DNA降解和代谢的研究进展[J].中国农业科技导报,2008,10(1):52-57.ZHANG X M,YU H M,WEI J,et al.Research progress on degradation and metabolism of foreign DNA in genetically modified food[J].Journal of Agricultural Science and Technology,2008,10(1):52-57.
[22]朱占齐,王卫国.转基因作物安全性及外源基因加工降解研究进展[J].饲料工业,2010,31(10):54-57.ZHU Z Q,WANG W G.Research advancement of gm crop safety and degradation of foreign gene[J].Feed Industry,2010,31(10):54-57.
[23]敖金霞.主要粮食作物深加工制品转基因检测技术研究[D].哈尔滨:东北农业大学,2008.AO J X.Study on transgenic detecting techniques for highly processed products of main crops[D].Harbin:Northeast Agricultural University,2008.
[24]柳方方,董美,李凯,等.转基因成分低水平混杂问题的现状[J].生物技术通报,2017,33(3):1-5.LIU F F,DONG M,LI K,et al.Global status on low level presence of genetically modified crops[J].Biotechnology Bulletin,2017,33(3):1-5.