Ontology type: schema:ScholarlyArticle
1989-05
AUTHORSMichael W. Lassner, Peter Peterson, John I. Yoder
ABSTRACTWe describe the simultaneous amplification of different segments of foreign DNA in transgenic plants using the polymerase chain reaction (PCR). We used PCR to simultaneously amplify different regions of transformed T-DNA in order to assay the integrity of transformed constructions in primary tomato transformants. We also used simultaneous PCR amplification to examine the segregation of transformed sequences in progeny of primary transformants. A tomato transformant containing the maize transposable elementAc was crossed to transformants containing the non-autonomousDs1 element flanked by maizeAdh1 sequences. We then ran PCR reactions on DNA from F1 progeny using two sets of primers, one set homologous toAc and one set homologous toAdh1 sequences on either side ofDs1. Because theAc andAdh1 primers resulted in amplification of fragments of different sizes, it was possible to monitor the inheritance ofAc and theDs1 containingAdh1 genein a single reaction. Additionally, it was possible to identify F1 plants in whichDs1 had excised by the amplification of a fragment the size predicted for an empty donor site. In order to run these reactions, we have constructed a simple and inexpensive thermal cycler which, when used in conjunction with the rapid miniscreen plant DNA isolation procedure described, allows the processing of a large number of samples in a single day. Therefore, we have shown that PCR can be a useful tool to monitor the integrity of foreign genes in transgenic plants, to follow the segregation of foreign DNA in progeny, and to assay for the excision of transposable elements. More... »
PAGES116-128
http://scigraph.springernature.com/pub.10.1007/bf02669627
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