A temperature-tolerant multiplex elements and genes screening system for genetically modified organisms based on dual priming oligonucleotide primers and capillary electrophoresis

Wei Fu; Shuang Wei; Chenguang Wang; Zhixin Du; Pengyu Zhu; Xiyang Wu; Gang Wu; Shuifang Zhu

doi:10.1016/j.foodchem.2017.02.088

A temperature-tolerant multiplex elements and genes screening system for genetically modified organisms based on dual priming oligonucleotide primers and capillary electrophoresis

Food Chem. 2017 Aug 15:229:396-402. doi: 10.1016/j.foodchem.2017.02.088. Epub 2017 Feb 21.

Authors

Wei Fu¹, Shuang Wei², Chenguang Wang³, Zhixin Du⁴, Pengyu Zhu¹, Xiyang Wu⁵, Gang Wu⁶, Shuifang Zhu⁷

Affiliations

¹ Institute of Plant Quarantine, Chinese Academy of Inspection and Quarantine, Beijing 100176, China.
² Shantou Entry-exit Inspection and Quarantine Bureau, Shantou 515041, China.
³ Institute of Plant Quarantine, Chinese Academy of Inspection and Quarantine, Beijing 100176, China; College of Plant Protection, China Agricultural University, Beijing 100193, China.
⁴ Guangxi Entry-exit Inspection and Quarantine Bureau, Nanning 530028, China.
⁵ Department of Food Science and Engineering, College of Science and Technology, Jinan University, Guangzhou 510632, China.
⁶ Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan 430062, China. Electronic address: wugang@caas.cn.
⁷ Institute of Plant Quarantine, Chinese Academy of Inspection and Quarantine, Beijing 100176, China. Electronic address: zhusf@caiq.gov.cn.

PMID: 28372191
DOI: 10.1016/j.foodchem.2017.02.088

Abstract

High throughput screening systems are the preferred solution to meet the urgent requirement of increasing number of genetically modified organisms (GMOs). In this study, we have successfully developed a multiplex GMO element screening system with dual priming oligonucleotide (DPO) primers. This system can detect the cauliflower mosaic virus 35S (CaMV 35S), terminator of nopaline synthase gene (NOS), figwort mosaic virus 35S (FMV 35S) promoter, neomycin phosphotransferaseII (NPTII), Bt Cry 1Ab, phosphinothricin acetyltransferase genes (bar) and Streptomyces viridochromogenes (pat) simultaneously, which covers more than 90% of all authorized GMO species worldwide. This system exhibits a high tolerance to annealing temperatures, high specificity and a limit of detection equal to conventional PCR. A total of 214 samples from markets, national entry-exit agencies, the Institute for Reference Materials and Measurement (IRMM) and the American Oil Chemists' Society (AOCS) were also tested for applicability. This screening system is therefore suitable for GMO screening.

Keywords: Capillary electrophoresis; DPO primers; GMO screening; High-throughput; Wide coverage.

MeSH terms

DNA Primers / genetics*
Electrophoresis, Capillary / methods*
Organisms, Genetically Modified / metabolism*
Promoter Regions, Genetic / genetics*

Substances

DNA Primers