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Acta Biochim Biophys Sin (Shanghai). 2013 May;45(5):416-21. doi: 10.1093/abbs/gmt016. Epub 2013 Feb 28.

Flanking sequence determination and event-specific detection of genetically modified wheat B73-6-1.

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1
Food Inspection Center, Liaoning Entry-Exit Inspection and Quarantine Bureau, Dalian 116001, China.

Abstract

In order to establish a specific identification method for genetically modified (GM) wheat, exogenous insert DNA and flanking sequence between exogenous fragment and recombinant chromosome of GM wheat B73-6-1 were successfully acquired by means of conventional polymerase chain reaction (PCR) and thermal asymmetric interlaced (TAIL)-PCR strategies. Newly acquired exogenous fragment covered the full-length sequence of transformed genes such as transformed plasmid and corresponding functional genes including marker uidA, herbicide-resistant bar, ubiquitin promoter, and high-molecular-weight gluten subunit. The flanking sequence between insert DNA revealed high similarity with Triticum turgidum A gene (GenBank: AY494981.1). A specific PCR detection method for GM wheat B73-6-1 was established on the basis of primers designed according to the flanking sequence. This specific PCR method was validated by GM wheat, GM corn, GM soybean, GM rice, and non-GM wheat. The specifically amplified target band was observed only in GM wheat B73-6-1. This method is of high specificity, high reproducibility, rapid identification, and excellent accuracy for the identification of GM wheat B73-6-1.

PMID:
23449073
DOI:
10.1093/abbs/gmt016
[Indexed for MEDLINE]
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