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  • Title: Detection of transgene copy number by analysis of the T1 generation of tobacco plants with introduced P3 gene of potato virus A.
    Author: Subr Z, Nováková S, Drahovská H.
    Journal: Acta Virol; 2006; 50(2):135-8. PubMed ID: 16808332.
    Abstract:
    Real-time PCR, namely the deltadeltaCt method was used to determine the relative copy number of Potato virus A (PVA) P3 gene in the genome of the T1 generation of 18 transgenic tobacco lines. These results were compared with segregation ratios of kanamycin (Km)-resistant phenotype in T1 plants of each line and were found to be, in general, concordant. All the five lines with the Mendelian segregation ratio of 3:1 carried one gene copy. In 12 of 13 lines with uneven segregation more inserted gene copies were detected. Only for one line the real-time PCR and phenotype segregation differed. According to our results the real-time PCR of T1 generation may be used as supplementary method of estimation of the number of transgene copies in the case of nonavailability of the original T0 plants.
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