|
|
|
| Plasmid pUC57-BT11 Construction and Application of Genetic Modified Maize Line BT11 |
| DONG Lian-hua,LI Liang,WANG Jing,ZHAO Zheng-yi,ZANG Chao |
| National Institute of Metrology, Beijing 100013, China |
|
|
|
|
Abstract To construct a plasmid pUC57-BT11 including event specific product and endogenous gene zSSIIb product.After restriction enzyme digestion and sequencing, the specific of the plasmid pUC57-BT11 is detected.The property value of pBT11 is 1.01±0.01(k=2) determined by sequencing and real time PCR.By comparison standard curve generated by plasmid with genomic DNA, no significant difference between the slopes of the standard curve is found.All correlated coefficient R2 are ≥0.99.Certified GM matrix materials are detected by quantitative PCR when pBT11 plasmid DNA is used as quantification standards.The results determined by quantitative PCR with pBT11 are identical with the certified values.This indicated that pBT11 plasmid can be used as a standard for GM maize BT11 quantitative and qualitatively detection.
|
|
|
|
|
|
|
|
［1］GB/T 19495.3—2004.转基因产品检测—核酸提取纯化方法［S］.
［2］GB/T 19495.4—2004.转基因产品检测—核酸定性PCR检测方法［S］.
［3］GB/T 19495.5—2004.转基因产品检测—核酸定量PCR检测方法［S］.
［4］Corbisier P, Broeders S, Charels D, et al.Certification of plasmidic DNA containing MON810 maize DNA fragments: certified reference materials ERM-AD413［R］.EUR 22948 EN,2007.
［5］Allnutt T R, Chisholm J, Hird H, et al. Plasmid standards for real time PCR and GM enforcement testing［EB］. http://www.gm-inspectorate.gov.uk/ documents/PLASMID.pdf,2011-02-05.
［6］Lee S,Park Y,Kim J,et al.Qualitative PCR method for detection of genetically modified maize lines NK603 and TC1507 ［J］.Agricul Chem Biotechnol, 2004, 47(4): 185-188.
［7］JCGM 100:2008. Evaluation of measurement data-guide to the expression of uncertainty in measurement［S］.
［8］ISO Guide 35:2006.Reference materials-General and statistical principles for certification［S］.
［9］Taverniers I, Windels P, Vaitilingom M, et al. Event-specific plasmid standards and real-time PCR methods for transgenic Bt11, Bt176, and GA21 maize and transgenic GT73 canola ［J］. J of Agricul Food Chem, 2005, 53(8): 3041-3052. |
|
|
|
2011, Vol. 32 
