Development and application of a general plasmid reference material for GMO screening

文献类型： 外文期刊

第一作者： Wu, Yuhua

作者： Wu, Yuhua;Li, Jun;Wang, Yulei;Li, Xiaofei;Li, Yunjing;Zhu, Li;Li, Jun;Wu, Gang;Wu, Yuhua;Li, Jun;Wang, Yulei;Li, Xiaofei;Li, Yunjing;Zhu, Li;Li, Jun;Wu, Gang

作者机构：

关键词： GMO;Screening;General positive control;Genetic element;Reference gene

期刊名称：PLASMID （ 影响因子：3.466； 五年影响因子：2.74 ）

ISSN： 0147-619X

年卷期： 2016 年 87-88 卷

页码：

收录情况： SCI

摘要： The use of analytical controls is essential when performing GMO detection through screening tests. Additionally, the presence of taxon-specific sequences is analyzed mostly for quality control during GMO detection. In this study, 11 commonly used genetic elements involving three promoters (P-35S, P-FMV35S and P-NOS), four marker genes (Bar, NPTII, HPT and Pmi), and four terminators (T-NOS, T-35S, T-g7 and T-e9), together with the reference gene fragments from six major crops of maize, soybean, rapeseed, rice, cotton and wheat, were co-integrated into the same single plasmid to construct a general reference plasmid pB1121-Screening. The suitability test of pBI121-Screening plasmid as reference material indicated that the non-target sequence on the pBI121-Screening plasmid did not affect the PCR amplification efficiencies of screening methods and taxon-specific methods. The sensitivity of screening and taxon-specific assays ranged from 5 to 10 copies of pBI121-Screening plasmid, meeting the sensitivity requirement of GMO detection. The construction of pBI121-Screening solves the lack of a general positive control for screening tests, thereby reducing the workload and cost of preparing a plurality of the positive control. (C) 2016 Elsevier Inc. All rights reserved.

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