转Cry1Ab基因水稻分子特征及其特异性PCR检测方法

doi:10.3724/SP.J.1005.2012.00208

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HEREDITAS ›› 2012, Vol. 34 ›› Issue (2): 208-214.doi: 10.3724/SP.J.1005.2012.00208

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Molecular characteristics and specific PCR detection of transgenic rice containing Cry1Ab

WANG Xiao-Fu¹, CHEN Xiao-Yun¹, ZHANG Xiao-Ming², ZHOU Yu¹, ZHANG Huan-Chun³, MIAO Qing-Mei¹, FANG Jing¹, XU Jun-Feng¹

1. Institute of Agricultural Quality and Standard, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China 2. Institute of Crop and Nuclear Technology Utilization, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China 3. College of Chemistry and Life Science, ShenYang Normal University, Shenyang 110034, China

Received:2011-05-24 Revised:2011-06-29 Online:2012-02-20 Published:2012-02-25

Abstract

Abstract: Bt01 is a new type of rice that has been genetically modified to express Cry1Ab protein. This study confirmed that Cry1Ab was inserted into Bt01 as a single copy using Southern blotting analysis. TAIL-PCR method was further used to obtain its insertion siteinformation. Specific PCR primers and TaqMan probes were designed based on the 5′-integration junction sequence of transgenic rice Bt01. The results showed that the limit of detection (LOD) was ten copies in qualitative PCR. The quantitative PCR assay showed that the LOD was five copies, and the limit of quantification (LOQ) was ten copies. In addition, the accuracy of the established quantitative PCR was verified by detecting two samples containing 3% and 0.5%Bt01, respectively. The quantitative PCRanalysis showed the results were 2.7% and 0.47%, respectively.The above results indicated that the event-specific PCR methodsdeveloped have high specificity and good sen-sitivity, which could be effective methods for identifying and testing the genetically modified Bt01 rice.

Key words: molecular characteristics, transgenic, Cry1Ab, qualitative and quantitative PCR, specific detection

WANG Xiao-Fu, CHEN Xiao-Yun, ZHANG Xiao-Ming, ZHOU Yu, ZHANG Huan-Chun, MIAO Qing-Mei, FANG Jing, XU Jun-Feng. Molecular characteristics and specific PCR detection of transgenic rice containing Cry1Ab[J]. HEREDITAS, 2012, 34(2): 208-214.

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Molecular characteristics and specific PCR detection of transgenic rice containing Cry1Ab

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