Qualitative detection and quantification of a <Emphasis Type="Italic">Cry1A(b)</Emphasis> transgene present in rice cv. Zhejing 22 |
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Authors: | Junfeng Xu Xiaofu Wang Xiaoyun Chen Xinquan Wang Yu Zhou Qingmei Miao Jing Fang |
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Affiliation: | (1) Institute of Agriculture Quality and Standard, Zhejiang Academy of Agricultural Sciences, Hangzhou, 310021, People’s Republic of China |
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Abstract: | 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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