抗环斑病毒转基因番木瓜551的PCR检测.doc

抗环斑病毒转基因番木瓜551的PCR检测 目录 TOC \o 1-9 \h \z \u 目录 1 正文 1 文1：抗环斑病毒转基因番木瓜551的PCR检测 1 1 材料与方法 3 13 DNA提取 3 15 PCR检测 4 2 结果 5 3 讨论 6 文2：抗环斑病毒转基因番木瓜551的PCR检测 7 1 材料与方法 8 13 DNA提取 9 15 PCR检测 10 2 结果 11 3 讨论 12 参考文摘引言： 13 原创性声明（模板） 14 文章致谢（模板） 15 正文 抗环斑病毒转基因番木瓜551的PCR检测 文1：抗环斑病毒转基因番木瓜551的PCR检测 PCRPCR for event-specific detection of tragenic virus resistant papaya 55-1 Abstract： Objective PCR assay was developed for event-specific detection of tragenic virus resistant papaya A developed Cetyl Trimethyl Ammonium bromide(CTAB) method and Qiagen DNeasy plant mini kit were used to extract DNA from papaya seed and prime for PCR detection of papaya 55-1 were designed to amplify endogenous papain gene sequence and external segments，35S-GUS and NOS-35S，and corresponding detection methods were Results of PCR amplification of papain gene indicated that both the developed CTAB method and Qiagen DNeasy plant mini kit were useful for DNA extraction from papaya seed and of amplification of the external segments (35S-GUS and NOS-35S) indicated that the developed method was useful to detect the tragenic virus resistant papaya line 55-1，and the absolute and relative limit of detection (LOD)of the method got to ×10-2 ng and % The developed PCR method can detect tragenic virus resistant papaya 55-1 specially and detective seitivity can satisfy the need of qualitative detection for genetically modified foods。 Key words： genetically modified;papaya 55-1;PCR 番木瓜属热带亚热带重要水果，为有效控制番木瓜环斑病毒(PV)对番木瓜产业的威胁，1998年世界上第1个转基因番木瓜品系――抗环斑病毒转基因番木瓜55-1在美国和加拿大获准商业种植〔1，2〕。从此转基因番木瓜55-1以其独特的抗病优势被广泛种植，每年都有大量转基因番木瓜流入国际水果市场。本研究采用改良十六烷基-三甲基-溴化铵(CTAB）法及试剂盒方法进行番木瓜基因组DNA提取，根据番木瓜管家Papain基因和抗环斑病毒转基因番木瓜55-1品系的外源结构基因(35S-GUS)和调控基因(NOS-35S)序列设计合成特异性检测引物，采用PCR技术，建立对抗环斑病毒转基因番木瓜55-1进行品系鉴定的检验方法，为今后转基因番木瓜的标识管理及安全性评价提供技术支持。 1 材料与方法 11 材料 转基因番木瓜55-1种籽由香港政府化验所友情提供。市场抽检番木瓜购自深圳市翠竹万佳超市及深圳市田贝一路街边水果店。 12 仪器和试剂 MJR PTC-220扩增仪，SynGene Gene Genius凝胶成像系统，Eppendorf Bio