抗环斑病毒转基因番木瓜55

抗环斑病毒转基因番木瓜 55 【关键词】 ,转基因；番木瓜 55-1； 摘要： 目的 建立对抗环斑病毒转基因番木瓜 55-1 进行品系鉴定的检测方法。方法 采纳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，采纳 PCR技术对抗环斑病毒转基因番木瓜 55-1 进行品系鉴定。结果 内源 Papain 基因 PCR 扩增结果表明，改良 CTAB 法及试剂盒方法都可用于番木瓜种籽及果肉的 DNA 提取。实验中合成的引物及建立的 PCR 反应体系、反应参数能特异性地扩增转基因番木瓜 55-1的外源基因 35S-GUS 序列和 NOS-35S 序列。该方法的绝对检测低限为 815×10-2ng，相对检测低限为 014％。结论 本检测方法有较高的稳定性，能有效地对转基因番木瓜55-1 进行品系鉴定，该方法的检测灵敏度可完全满足转基因食品标识管理定性检测的需要。 关键词： 转基因；番木瓜 55-1； PCRPCR for event-specific detection of transgenic virus resistant papaya 55-1 Abstract： Objective PCR assay was developed for event-specific detection of transgenic 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 primers 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 transgenic 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 d...