Barnase基因定性PCR检测方法及其标准化  

Development and Standardization of Qualitative PCR Detection Method Targeting barnase Gene

作　　者：李俊[1] 沈旻伟 武玉花[1] 吴刚[1] 

机构地区：[1]中国农业科学研究院油料作物研究所农业部油料作物生物学与遗传育种重点实验室,武汉430062

出　　处：《生物技术通报》2013年第12期129-136,共8页Biotechnology Bulletin

基　　金：国家自然科学基金项目(31271810);国家转基因生物新品种培育重大专项(2013ZX08012-003)

摘　　要：Barnase基因编码核糖核酸酶,在基因工程育种中通常被用于创建雄性不育系。为了满足转barnase基因作物安全监管的需要,建立了barnase基因普通PCR、实时荧光PCR检测方法,具有良好的扩增特异性和检测灵敏度。barnase基因定性PCR检测方法经过了国内8家有资质的实验室的循环验证,结果表明该方法能够特异性检测样品中barnase基因,检测灵敏度均达到0.10%以上,且检测结果具有良好的重复性和重现性。符合标准方法检测特异性强、灵敏度高、重复性好的要求,为我国转barnase基因作物检测和转基因生物安全管理制度的实施提供了相应的技术支撑。Barnase gene, encoding a ribonuclease, is usually used for developing male sterility line in genetic engineering research. In order to meet the requirements of safety regulation of transgenic crops, the conventional PCR and real-time PCR detection methods targeting the Barnase gene were developed, displaying exollent amplification specificity and test sensitivity. The qualitative methods targeting Barnase gene were validated by 8 certified laboratories, results showed that this method could specifically detect the Barnase gene from testing samples with satisfactory detection sensitivity of about 0.1%, fine repeatability and reproducibility, meeting the performance criteria of standard methods. This study established a suitable method for detection of the transgenic crops harboring the Barnase gene, providing technical support for the safety management of transgenic crops in China.

关 键 词：转基因检测 Barnase基因 普通PCR 实时荧光PCR 循环验证 

分 类 号：Q943.2[生物学—植物学]