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dc.contributorIraqi, Drissen_US
dc.contributorUdupa, Sripada M.en_US
dc.contributorEl Alaoui, My Abdelazizen_US
dc.contributorAlaoui, Sanaaen_US
dc.contributorIbriz, Mohammeden_US
dc.contributorElfahime, Elmostafaen_US
dc.creatorMelloul, Marouaneen_US
dc.date2014-01-18en_US
dc.date.accessioned2017-07-24T01:01:17Z
dc.date.available2017-07-24T01:01:17Z
dc.identifierhttps://mel.cgiar.org/reporting/download/hash/zHKlXlABen_US
dc.identifier.citationMarouane Melloul, Driss Iraqi, Sripada M. Udupa, My Abdelaziz El Alaoui, Sanaa Alaoui, Mohammed Ibriz, Elmostafa Elfahime. (18/1/2014). Development of specific primers for the detection of HVA1 from barley in transgenic durum wheat by polymerase chain reaction (PCR) technology. African journal of biotechnology, 13 (4), pp. 581-592.en_US
dc.identifier.urihttps://hdl.handle.net/20.500.11766/7295
dc.description.abstractGenetic transformation is a widely employed tool in both basic research and commercial plant breeding programs. Its application requires that transgenes be stably integrated and expressed in the plant genome. When transgenic plants are developed, it is essential to determine which plants contain the transgene. Detection methods are usually based on amplification of the target transgene. This paper describes a development of detection method based on conventional and real time polymerase chain reaction (PCR) for simultaneous detection of barley HVA1 transgene and its transcript in transformed durum wheat. Since there exist a high homology between the barley HVA1 gene and the wheat gene, development of a specific sets of primers is needed for PCR-based characterizations, and the study of the transgene. Based on the alignment of the two genes sequences obtained from public databases, several primers were designed to detect and distinguish between the transformed and non-transformed plants. Real time PCR has been employed because of its inherent sensitivity and quantitative nature. It has been possible to design the following primers pairs F2/MMR, F2/R10 and F14/R10 as highly specific and suitable for the detection of HVA1 DNA by conventional and real-time PCR. Nonetheless, the primers used were allowed to reach high efficiencies and did not show any cross-reactivity with DNAs extracted from various plants. The sensitivity achieved was 6.4 pg. The primer pair F2/R10 was considered as highly specific for the detection of both DNA and mRNA of the HVA1 by real-time PCR. The assays proved to be accurate, specific, sensitive and sufficiently reproducible for further application in high-throughput molecular characterization of transgenic lines.en_US
dc.formatPDFen_US
dc.languageenen_US
dc.publisherAcademic Journalsen_US
dc.rightsCC-BY-NC-4.0en_US
dc.sourceAfrican journal of biotechnology;13,(2014) Pagination 581,592,en_US
dc.subjecthva1en_US
dc.subjectreal time polymerase chain reaction (pcr)en_US
dc.subjecttransgenic planten_US
dc.subjectBarleyen_US
dc.subjectWheaten_US
dc.titleDevelopment of specific primers for the detection of HVA1 from barley in transgenic durum wheat by polymerase chain reaction (PCR) technologyen_US
dc.typeJournal Articleen_US
cg.creator.idUdupa, Sripada M.: 0000-0003-4225-7843en_US
cg.creator.ID-typeORCIDen_US
cg.subject.agrovocdroughten_US
cg.subject.agrovocdurum (triticum durum)en_US
cg.contributor.centerIbn Tofail University - UIT Moroccoen_US
cg.contributor.centerNational Institute of Agronomic Research Morocco - INRA Moroccoen_US
cg.contributor.centerInternational Center for Agricultural Research in the Dry Areas - ICARDAen_US
cg.contributor.centerNational Center for Scientific and Technical Research - CNRSTen_US
cg.contributor.centerThe French National Center for Scientific Research - CNRSen_US
cg.contributor.centerIbn Tofail University, Faculty of Sciences - UIT - FSen_US
cg.contributor.crpCGIAR Research Program on Dryland Systems - DSen_US
cg.contributor.funderInternational Fund for Agricultural Development - IFADen_US
cg.contributor.projectEnhanced small-holder wheat-legume cropping systems to improve food security under changing climate in the drylands of West Asia and North Africaen_US
cg.contributor.project-lead-instituteInternational Center for Agricultural Research in the Dry Areas - ICARDAen_US
cg.coverage.regionNorthern Africaen_US
cg.coverage.countryMAen_US
cg.contactelfahime@cnrst.maen_US
cg.identifier.doihttps://dx.doi.org/10.5897/AJB2013.13301en_US
dc.identifier.statusOpen accessen_US
mel.project.openhttps://mel.cgiar.org/projects/46en_US


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