Integrated structure and event-specific real-time detection of transgenic cry1Ac/SCK rice Kefeng 6
| العنوان: | Integrated structure and event-specific real-time detection of transgenic cry1Ac/SCK rice Kefeng 6 |
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| المؤلفون: | Xifeng Wang, Changqing Su, Jiajian Xie, Yufa Peng |
| المصدر: | European Food Research and Technology. 232:351-359 |
| بيانات النشر: | Springer Science and Business Media LLC, 2010. |
| سنة النشر: | 2010 |
| مصطلحات موضوعية: | Genetics, Transgene, General Chemistry, Biology, Biochemistry, Genome, Genetically modified rice, Industrial and Manufacturing Engineering, Transformation (genetics), Real-time polymerase chain reaction, Plasmid, Primer walking, Gene, Food Science, Biotechnology |
| الوصف: | Transgenic rice Kefeng 6 is a transformation event containing two insect-resistant genes, cry1Ac and SCK (modified CpTI gene) in China. In order to monitor the probable release of Kefeng 6 in the future and execute the labeling requirements, it is necessary to develop a rapid and reliable detection method. In this study, both the 5′ and 3′-junction sequences spanning the plant DNA and the integrated gene construct of the rice event Kefeng 6 were isolated by genome walking and long-distance PCR (LD-PCR), successively. Multiple copies of truncated SCK gene and cry1Ac gene were found to integrate into the host rice genome. The event-specific real-time detection method for Kefeng 6 event based on its 5′-junction sequence was established using one plasmid molecule pMD-KF6 containing both 5′-junction sequence and rice endogenous gene gos9 sequence as the reference material (RM) with an absolute limit of quantification (LOQa) around 10 template copies. Thereafter, three different transgenic amounts of w/w mixed samples (5, 1, and 0.5%, respectively) were quantified to assess the performance characteristics of the established real-time PCR method. The accuracy expressed as bias deviated from the 4.00–26.00%, the precision expressed as standard deviation (SD) and relative standard deviation (RSD) deviated from 0.03–0.19 and 3.42–4.76%, respectively. Based on the earlier results, we concluded that the qualitative and quantitative PCR assays were reliable and accurate for Kefeng 6 measurement, and the reference plasmid pMD-KF6 could be a good substitute for the reference material for Kefeng 6 quantification. |
| تدمد: | 1438-2385 1438-2377 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_________::5dea93fee1fc6165b864f42d818610ad https://doi.org/10.1007/s00217-010-1399-z |
| حقوق: | CLOSED |
| رقم الأكسشن: | edsair.doi...........5dea93fee1fc6165b864f42d818610ad |
| قاعدة البيانات: | OpenAIRE |
Find this article in full text from Springer Verlag. | تدمد: | 14382385 14382377 |
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