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dc.creatorFlachowsky,Henryk
dc.creatorRiedel,Marko
dc.creatorReim,Stefanie
dc.creatorHanke,Magda-Viola
dc.date2008-01-01
dc.date.accessioned2019-05-03T12:44:28Z
dc.date.available2019-05-03T12:44:28Z
dc.identifierhttps://scielo.conicyt.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000100003
dc.identifier.urihttp://revistaschilenas.uchile.cl/handle/2250/84988
dc.descriptionThe generation of transgenic apple plants relies on the molecular analysis of transgene integration and expression based on polymerase chain reaction (PCR) analysis, blotting techniques and enzymatic assays on vitro leaves of putative transgenic regenerates. In order to assess the uniformity and the stability of transfer DNA (T-DNA) integration and gene expression, we studied 26 transgenic apple lines carrying the attacin E gene from Hyalophora cecropia, the β-glucuronidase gene, and the nptII gene. Plants were evaluated using standard molecular techniques, such as PCR, Southern blot, reverse transcription PCR (RT-PCR) and Enzyme Linked Immunosorbent Assay (ELISA), and propagated in vitro on non-selective antibiotic-free media for four years to mimic natural conditions in the field. In some T-lines transgene integration and expression did not remain stable; differences were also found between distinct plants of a single T-line. Individual plants with partially or completely silenced transgenes were identified as well as plants with non-detectable T-DNA. Several lines appeared chimeric or partially silenced. Although most molecular techniques can reliably detect the presence of transgenic cells, they often fail to detect mixtures of transformed and non-transformed cells, or cells with silenced transgenes. This should be taken into consideration, especially in the case of vegetatively propagated trees, where non-transformed or silenced plant parts could mistakenly be used as propagation material.
dc.formattext/html
dc.languageen
dc.publisherPontificia Universidad Católica de Valparaíso
dc.relation10.4067/S0717-34582008000100003
dc.rightsinfo:eu-repo/semantics/openAccess
dc.sourceElectronic Journal of Biotechnology v.11 n.1 2008
dc.subjectAgrobacterium tumefaciens
dc.subjectapple
dc.subjectchimeric tissue
dc.subjectgene silencing
dc.subjectMalus domestica
dc.subjectT-DNA leakage
dc.titleEvaluation of the uniformity and stability of T-DNA integration and gene expression in transgenic apple plants


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