Qualitative detection and quantification of a Cry1A(b) transgene present in rice cv. Zhejing 22

文献类型： 外文期刊

第一作者： Xu, Junfeng

作者： Xu, Junfeng;Wang, Xiaofu;Chen, Xiaoyun;Wang, Xinquan;Zhou, Yu;Miao, Qingmei;Fang, Jing

作者机构：

关键词： Qualitative and quantitative PCR;GMOs;Event-specific detection;Bt-ZJ22;TAIL-PCR;Transgene integration

期刊名称：EUROPEAN FOOD RESEARCH AND TECHNOLOGY （ 影响因子：2.998； 五年影响因子：3.005 ）

ISSN： 1438-2377

年卷期： 2011 年 233 卷 2 期

页码：

收录情况： SCI

摘要： The rice cultivar Zhejing 22 has been transformed to carry the Bacillus thuringiensis Cry1A(b) gene in 2007. Although the cultivation of this line (referred to as Bt-ZJ22) is yet to be authorized, it is likely to be approved in China in the near future. The transgene-host 3' integration junction was isolated using thermal asymmetric interlaced PCR, and its sequence was used to design PCR primers and an appropriate TaqMan probe for a real-time PCR quantitative assay of the transgene. The limit of detection for a standard PCR assay was estimated at ten gene copies, while that for the real-time PCR assay was 5-10 copies. The latter assay was tested on samples of rice DNA with either 5, 3, 1, or 0.5% Bt-ZJ22; this produced estimates for the relative content of Bt-ZJ22 of, respectively, 5.3, 3.2, 0.97, and 0.48%. The precision of these estimates demonstrated that this real-time PCR assay should be suitable for the detection and monitoring of this transgenic event.

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