转基因玉米TC1507质粒DNA标准物质的研制

标准物质具有特定量值、均匀性和稳定性三大典型特征,也是其作为测量标尺的依据。转基因生物标准物质是我国转基因产品标识制度顺利实施的关键技术支撑之一。文章以转基因玉米TC1507为对象,制备了转化体特异性的新型质粒DNA标准物质pTC1507,并对其均匀性、稳定性、量值进行了评价和测定。测试结果表明,制备的质粒DNA标准物质具有良好的瓶间和瓶内均匀性,pTC1507稳定性可靠,可以在-20 ℃稳定放置6个月以上。经过测序和实时荧光定量PCR定值以及不确定度评估,pTC1507标准物质的量值是1.01±0.053。均匀性、稳定性和量值结果表明,研制的转基因玉米TC1507质粒分子标准物质符合标准物质的典型要求,可以代替传统的基体标准物质应用于转基因玉米检测,解决传统标准物质获取困难、制备复杂、成本高等不足。

Development of plasmid DNA reference material of genetically modified maize TC1507

The reference material has three typical characteristics of characterization value, homogeneity and stability, which was used as the scale of biological measurement. The development of reference materials for genetically modified organisms was one of the key technologies for the implementation of the GMO labeling system in China. In this study, a novel plasmid DNA standard substance pTC1507 specific to the transgene maize TC1507 was produced, and its homogeneity, stability and characterization were evaluated and measured. The results showed that the pTC1507 had good homogeneity between and within bottles. The stability of pTC1507 was reliable, and it could be stably placed at -20 ℃ for 6 months at least. After sequencing, quantitative real-time PCR analysis, and uncertainty evaluation, the characterization value of pTC1507 was 1.01±0.053. The results of homogeneity, stability and characterization evaluation showed that pTC1507 was satisfied with the typical requirements of reference materials and could replace the conventional matrix-based reference materials in the detection of transgenic maize. The developed plasmid reference material showed the advantages of easy producing, simple preparation, and low cost.