| 玉米转基因成分的非同源模板竞争定量PCR检测 |
| |
| 引用本文： | 栾春光,马林,郝彦玲,朱本忠,罗云波.玉米转基因成分的非同源模板竞争定量PCR检测[J].河南工业大学学报(自然科学版),2004,25(1):19-21. |
| |
| 作者姓名： | 栾春光 马林 郝彦玲 朱本忠 罗云波 |
| |
| 作者单位： | 中国农业大学,食品科学与营养工程学院,北京,100083 |
| |
| 摘 要： | 建立了玉米转基因成分中35S启动子非同源模板的竞争定量PCR检测系统.竞争定量PCR的关键问题是内标物的制备.实验中非同源模板的构建是采用PCR方法对大肠杆菌基因组进行低严紧型扩增,回收预期DNA片段并将其构建到T-easy载体上.通过调整非同源模板浓度使竞争物PCR产物亮度与1%GMO玉米的亮度相同,从而确定转基因玉米的检出限为1%.然后以确定的内标物浓度与不同含量的GMO玉米样品进行竞争定量PCR反应,以此进一步确定样品中GMO的含量范围.
|
| 关 键 词： | 竞争定量PCR 非同源模板 35S启动子 转基因玉米 |
| 文章编号： | 1671-1629(2004)01-0019-03 |
| 修稿时间： | 2003年12月1日 |
HETEROLOGOUSLY QUANTITATIVE COMPETITIVE PCR FOR THE DETECTION OF GENETICALLY MODIFIED MAIZE |
| |
| LUAN Chun-guang,MA Lin,HAO Yan-ling,ZHU Ben-zhong,LUO Yun-bo.HETEROLOGOUSLY QUANTITATIVE COMPETITIVE PCR FOR THE DETECTION OF GENETICALLY MODIFIED MAIZE[J].Journal of Henan University of Technology Natural Science Edition,2004,25(1):19-21. |
| |
| Authors: | LUAN Chun-guang MA Lin HAO Yan-ling ZHU Ben-zhong LUO Yun-bo |
| |
| Abstract: | The quantitative competitive polymerase chain reaction (QC-PCR) system of using 35S promoter heterologous template for the detection of genetically modified maize was developed in this study.The key problem of QC-PCR is the construction of internal standard.The heterologous template is constucted as follow:the E.coli genomic DNA was low stringently amplified,the selected band was purified and ligated pGEM-T Easy Vector.The competitor concentrations were adjusted by the content of the heterologous template in a way that the equivalence point represented a GMO content of 1%.Therefore,the minimum detectable quantity was 1%.Then template DNA mixtures containing different proportion GMO were co-amplified with constant internal standard concentration of further determine GMO content in food samples. |
| |
| Keywords: | competitive quantitative PCR heterologous template 35S promoter genetically modified maize |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
