转基因溶菌酶羊hLZ-J1品系特异性数字PCR方法及样本中痕量转基因成分检测  

Development of an Event-Specific Droplet Digital PCR Assay for Quantification of the Transgene DNAs in Trace Samples of GM Goat hLZ-J1

作　　者：王艺洁 徐文婷 杨立桃[1] 梁晋刚 WANG Yijie;XU Wenting;YANG Litao;LIANG Jingang(School of Life Sciences and Biotechnology,Shanghai Jiao Tong University,Shanghai 200240,China;Development Center of Science and Technology,Ministry of Agriculture of People’s Republic of China,Beijing 100176,China)

机构地区：[1]上海交通大学生命科学技术学院 [2]农业农村部科技发展中心

出　　处：《中国生物防治学报》2022年第5期1109-1116,共8页Chinese Journal of Biological Control

基　　金：转基因重大专项(2016ZX08012-005);农业科研杰出人才培养计划(2021)。

摘　　要：人溶菌酶是一种天然抗生素,具有抗细菌、真菌,抗炎症,预防感染等功能。转基因溶菌酶山羊可以在乳腺中特异性表达,从而可以从羊乳中规模化制备溶菌酶。本研究旨在建立一种适用于痕量样本中转基因溶菌酶羊成分定量检测的品系特异性数字PCR方法,并利用该方法对山羊的血液、羊奶、粪便和环境土壤样本等进行检测,用于对转基因溶菌酶山羊成分的溯源。建立的数字PCR方法具有高特异性、灵敏度、准确性,检测限和定量限低至15个拷贝每个反应,可以对羊血、羊乳等制品进行准确的定量检测。研究结果还表明该方法可以准确测定粪便和环境土壤样本中的痕量转基因溶菌酶羊成分,可进一步用于转基因动物外源基因的水平转移检测及环境风险评估。Human lysozyme(hLZ) is a natural antibiotic,functioning to resist bacteria,fungi,inflammation and infection.Transgenic human lysozyme goat can express high levels of human lysozyme in milk for large scale production of lysozyme.Herein,we developed an event-specific droplet digital PCR(ddPCR) method for quantitative detection of transgenic hLZ goat and its derivates.This method presents high specificity,sensitivity and accuracy in goat blood and goat milk samples analysis.The limit of detection and limit of quantification were determined with 15 copies per reaction.Furthermore,this method was used to trace the transgenic hLZ goat content in goat feces and the living environmental soil samples,indicating that the developed ddPCR method can be used for monitoring horizontal gene transfer and environmental risk assessment of transgenic hLZ goat.

关 键 词：数字PCR 转基因溶菌酶羊 品系特异性 痕量样本 

分 类 号：S476[农业科学—农业昆虫与害虫防治]