以实时荧光定量PCR 技术检测转基因玉米MON8801.pdfVIP

作物学报 ACTA AGRONOMICA SINICA 2011, 37(11): 21172121 /zwxb/ ISSN 0496-3490; CODEN TSHPA9 E-mail: xbzw@ DOI: 10.3724/SP.J.1006.2011.02117 以实时荧光定量PCR 技术检测转基因玉米MON88017 袁 磊1,2 孙红炜1 李 凡1 李 宁1,3 赵 蕾3,* 路兴波1,* 1 / , 250100; 2 , 264670; 3 , 250014 : MON88017 (zSSIIb) Taqman , PCR MON88017 , 5 MON88017 (0.01%0.05%0.10%0.50% 1.00%) , 19~30 PCR : ; PCR; MON88017; Detection of Genetically Modified Maize MON88017 by Quantitative Real- time PCR 1,2 1 1 1,3 3,* 1,* YUAN Lei , SUN Hong-Wei , LI Fan , LI Ning , ZHAO Lei , and LU Xing-Bo 1 Institute of Plant Protection, Shandong Academy of Agricultural Sciences, Ji’nan 250100, China; 2 Department of Food Engineering, Shandong Business Institute, Yantai 264670, China; 3 College of Life Science, Shandong Normal University, Ji’nan 250014, China Abstract: Transgenic event-specific primers and Taqman probes based on the left-flanking sequence of MON88017 and endoge- nous gene (zSSIIb) were designed, and a real-time PCR method was developed to quantitatively detect the event-specific geneti- cally modified maize. The reference molecule composed of endogenous gene and flanking sequence of MON88017 was con- structed artificially. Two standard curves of the reference gene sequence and the 5 flanking sequence were established. We de- tected five mixed samples, and their genetically modified contents of MON88017 were 0.01%, 0.05%, 0.1%, 0.5%, and 1%, re- spectively. The lowest detection limit was 19–30 copies. This study indicated that the Taqman probes real-time PCR is highly specific and sensitive. Keywords: Gene