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Acta Agron Sin ›› 2013, Vol. 39 ›› Issue (08): 1501-1506.doi: 10.3724/SP.J.1006.2013.01501

• RESEARCH NOTES • Previous Articles     Next Articles

Construction and Application of a Reference Plasmid Molecule Suitable for phyA2 of Phytase Transgenic Maize

ZHANG Guang-Yuan1,2,SUN Hong-Wei1,LI Fan1,YANG Shu-Ke1,LU Xing-Bo1,*,ZHAO Lei2,*   

  1. 1 Institute of Plant Protection, Shandong Academy of Agricultural Sciences, Shandong Key Laboratory of Plant Virology, Jinan 250100, China; 2 Shandong Normal University, Jinan 250014, China
  • Received:2013-01-21 Revised:2013-04-22 Online:2013-08-12 Published:2013-05-20
  • Contact: 路兴波,E-mail: luxb99@sina.com, Tel: 0531-83179095;赵蕾,E-mail: zhaolei@sdu.edu.cn,Tel: 0531-88177190

Abstract:

Plasmid molecule based reference material has been shown to be a good alternative as the calibrator for genetically modified organisms (GMOs) identification and quantification. In this study, primers and probe for qualitative and quantitative PCR detection for phytase transgenic maize were designed, and a flexible plasmid pPZ containing phyA2 (243 bp) and zSSIIb (178 bp) sequences, was developed. Primers and plasmid were specifically tested through qualitative PCRs. Results showed that the primers and pPZ plasmid were very specific for phytase transgenic maize since there were no other PCR products amplified using different GMOs and primers except the specific main bands. Four different levels of genetically modified (GM) matrix samples (3.0%, 1.0%, 0.5%, and 0.1%) were detected by real-time PCR when plasmid pPZ and phytase transgenic maize genomic DNA were used as quantification standards, respectively. t-test showed the slop, linearity (R2) and quantification of the two standards for phyA2 and zSSIIb were not significantly different. The results indicated that the pPZ plasmid can be used as standard for phytase transgenic maize detection.

Key words: Phytase transgenic maize, Plasmid, Taqman

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