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Plasmid. 2016 Sep - Nov;87-88:28-36. doi: 10.1016/j.plasmid.2016.08.001. Epub 2016 Aug 4.

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

Author information

1
Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, No. 2 Xudong 2nd Road, Wuhan 430062, China; Supervision and Test Center (Wuhan) for Environmental Safety of Genetically Modified Plants, Ministry of Agriculture, No. 2 Xudong 2nd Road, Wuhan 430062, China.
2
Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, No. 2 Xudong 2nd Road, Wuhan 430062, China; Supervision and Test Center (Wuhan) for Environmental Safety of Genetically Modified Plants, Ministry of Agriculture, No. 2 Xudong 2nd Road, Wuhan 430062, China. Electronic address: wugang@caas.cn.

Abstract

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 pBI121-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.

KEYWORDS:

GMO; General positive control; Genetic element; Reference gene; Screening

PMID:
27497661
DOI:
10.1016/j.plasmid.2016.08.001
[Indexed for MEDLINE]

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