转植酸酶基因玉米phya2基因标准质粒分子构建及其检测-作物学报.pdfVIP

作物学报 ACTA AGRONOMICA SINICA 2013, 39(8): 1501−1506 /zwxb/ ISSN 0496-3490; CODEN TSHPA9 E-mail: xbzw@ DOI: 10.3724/SP.J.1006.2013.01501 转植酸酶基因玉米phyA2 基因标准质粒分子构建及其 检测应用 张广远1,2 孙红炜1 李 凡1 杨淑珂1 路兴波1,* 赵 蕾2,* 1 / , 250100; 2 , 250014 : phyA2 , phyA2 (243 bp)(178 bp)pPZ DNA phyA2 , PCR 131 bp ; pPZ, pPZ DNA (131 bp 88 bp)pPZ DNA 4 (3.0%1.0%0.5%0.1%) , t , 2 , pPZ : ; ; TaqMan Construction and Application of a Reference Plasmid Molecule Suitable for phyA2 of Phytase Transgenic Maize 1,2 1 1 1 1,* 2,* ZHANG Guang-Yuan , SUN Hong-Wei , LI Fan , YANG Shu-Ke , LU Xing-Bo , and ZHAO Lei 1 Institute of Plant Protection, Shandong Academy of Agricultural Sciences, Shandong Key Laboratory of Plant Virology, Jinan 250100, China; 2 College of Life Science, Shandong Normal University, Jinan 250014, China 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 pr