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Detection and identification of genetically modified EE-1 brinjal (Solanum melongena) by single, multiplex and SYBR real-time PCR.
Rajashekhar, V. Ballari and Asha, Martin and Gowda, Lalitha R. (2013) Detection and identification of genetically modified EE-1 brinjal (Solanum melongena) by single, multiplex and SYBR real-time PCR. Journal of the Science of Food and Agriculture, 93 (2). pp. 340-347. ISSN 0022-5142
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Abstract
BACKGROUND: Brinjal is an important vegetable crop. Major crop loss of brinjal is due to insect attack. Insect-resistant EE-1 brinjal has been developed and is awaiting approval for commercial release. Consumer health concerns and implementation of international labelling legislation demand reliable analytical detectionmethods for genetically modified (GM) varieties. RESULTS: End-point and real-time polymerase chain reaction (PCR) methods were used to detect EE-1 brinjal. In end-point PCR, primer pairs specific to 35S CaMV promoter, NOS terminator and nptII gene common to other GM crops were used. Based on the revealed 3� transgene integration sequence, primers specific for the event EE-1 brinjal were designed. These primers were used for end-point single, multiplex and SYBR-based real-time PCR. End-point single PCR showed that the designed primers were highly specific to event EE-1 with a sensitivity of 20 pg of genomic DNA, corresponding to 20 copies of haploid EE-1 brinjal genomic DNA. The limits of detection and quantification for SYBR-based real-time PCR assay were 10 and 100 copies respectively. CONCLUSION: The prior development of detection methods for this important vegetable crop will facilitate compliance with any forthcoming labelling regulations.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | genetically modified organisms; cry1Ac gene; event-specific PCR; Bt brinjal |
| Subjects: | 500 Natural Sciences and Mathematics > 07 Life Sciences > 03 Biochemistry & Molecular Biology 600 Technology > 08 Food technology > 23 Vegetables |
| Divisions: | Food Safety Analytical Quality Control Lab |
| Depositing User: | Food Sci. & Technol. Information Services |
| Date Deposited: | 16 Nov 2012 10:44 |
| Last Modified: | 28 Jun 2016 08:27 |
| URI: | http://ir.cftri.res.in/id/eprint/11019 |
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