| dc.creator | Hadi,Faranak | |
| dc.creator | Salmanian,Ali Hatef | |
| dc.creator | Ghazizadeh,Elham | |
| dc.creator | Amani,Jafar | |
| dc.creator | Noghabi,Kambiz Akbari | |
| dc.creator | Mousavi,Amir | |
| dc.date | 2012-07-01 | |
| dc.date.accessioned | 2019-05-03T12:44:58Z | |
| dc.date.available | 2019-05-03T12:44:58Z | |
| dc.identifier | https://scielo.conicyt.cl/scielo.php?script=sci_arttext&pid=S0717-34582012000400002 | |
| dc.identifier.uri | http://revistaschilenas.uchile.cl/handle/2250/85279 | |
| dc.description | Background: For successful in vitro plant regeneration, plant cell lines with multiple transgene integration and low transgene expression levels need to be ruled out. Although real-time polymerase chain reaction (RT-PCR) is a rapid way to accomplish this, it is also expensive and typically limits the size of the target sequence. Quantitative competitive PCR (QC-PCR) is proven to be a safe and accurate method for determination of both copy number and quantification of transcript levels of synthetic transgenes in transformed plants. Results: The glyphosate oxidoreductase genewas chemically synthesized and used to transform Brassica napus L. via Agrobactrium-mediated transformation. A construct containing the mutated form of a synthetic glyphosate oxidoreductase (gox) gene (internal standard) was prepared. Gene copy number was estimated in nine independent transgenic lines using QC-PCR as well as the standard method of Southern blot analysis. By quantitative RT-PCR, transcript levels were also determined in these lines. High (> 3), medium to high (2.2-3), medium to low (1-2.2), and low (< 1) levels of transcript were detected. Conclusions: No direct relationship was found between copy number and transgene expression levels. QC-PCR method could be implemented to screen putative transgenic plants and quickly select single T-DNA inserts. QC-PCR methods and the prepared competitor construct may be useful for future quantification of commercial transgenic food and feed. | |
| dc.format | text/html | |
| dc.language | en | |
| dc.publisher | Pontificia Universidad Católica de Valparaíso | |
| dc.rights | info:eu-repo/semantics/openAccess | |
| dc.source | Electronic Journal of Biotechnology v.15 n.4 2012 | |
| dc.subject | Brassica napus L. | |
| dc.subject | competitive quantitative PCR | |
| dc.subject | transcript level | |
| dc.subject | transgene copy number | |
| dc.title | Development of quantitative competitive PCR for determination of copy number and expression level of the synthetic glyphosate oxidoreductase gene in transgenic canola plants | |
