Event-specific detection of genetically modified wheat B73-6-1 based on the 3′-flanking sequence
In this study, 3′-flanking sequence between the host plant DNA and the integrated gene construct of pHMW1Dx5 vector in transgenic wheat B73-6-1 was revealed by means of adaptor PCR; thus, the fragment with the length of 3.1 kb was obtained, including a 190-bp wheat genomic DNA, which demonstrates th...
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Published in | European food research & technology Vol. 235; no. 6; pp. 1149 - 1159 |
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Main Authors | , , , , , , , |
Format | Journal Article |
Language | English German |
Published |
Berlin/Heidelberg
Springer-Verlag
01.12.2012
Springer Springer Nature B.V |
Subjects | |
Online Access | Get full text |
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Summary: | In this study, 3′-flanking sequence between the host plant DNA and the integrated gene construct of pHMW1Dx5 vector in transgenic wheat B73-6-1 was revealed by means of adaptor PCR; thus, the fragment with the length of 3.1 kb was obtained, including a 190-bp wheat genomic DNA, which demonstrates that this HMW-GS gene was located on the wheat chromosome 3B. And the event-specific PCR primers were designed based upon the revealed 3′-flanking sequence; the conventional qualitative PCR and quantitative SYBR real-time PCR detection methods employing these primers were successfully developed. In conventional qualitative PCR assay, the limit of detection was 0.1 % for B73-6-1 wheat genomic DNA for one reaction. In the quantitative SYBR real-time PCR assay, the limit of detection and limit of quantification were 10 and 100 haploid genome copies, respectively. In addition, three mixed blind wheat samples with known B73-6-1 contents were detected using the established real-time PCR systems, and the ideal results indicated that the established event-specific real-time PCR detection systems were reliable, sensitive and accurate. |
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Bibliography: | ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 |
ISSN: | 1438-2377 1438-2385 |
DOI: | 10.1007/s00217-012-1848-y |