Detection of transgene copy number by analysis of the T1 generation of tobacco plants with introduced P3 gene of potato virus A

Acta Virologica

Volumn 50, Issue 2, 2006, Pages 135-138

  Subr Z ^a   Novakova S ^a   Drahovska H ^b  

^a INSTITUTE OF VIROLOGY   (Slovakia)

^b Faculty of Natural Sciences   (Slovakia)

Author keywords

Phenotypic segregation; Potato virus A; Potyvirus; Real time PCR; Transgenic tobacco

Indexed keywords

KANAMYCIN; VIRUS DNA; VIRUS PROTEIN;

ANTIBIOTIC RESISTANCE; ARTICLE; CONTROLLED STUDY; GENE NUMBER; NONHUMAN; NUCLEOTIDE SEQUENCE; P3 GENE; PHENOTYPE; PLANT VIRUS; POTATO VIRUS A; REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION; TOBACCO; TRANSGENE; VIRUS GENE; GENE DOSAGE; GENETICS; METHODOLOGY; POLYMERASE CHAIN REACTION; POTYVIRUS; TRANSGENIC PLANT;

NICOTIANA OBTUSIFOLIA; NICOTIANA TABACUM; POTATO VIRUS A; POTYVIRUS;

DNA, VIRAL; GENE DOSAGE; GENES, VIRAL; PLANTS, GENETICALLY MODIFIED; POLYMERASE CHAIN REACTION; POTYVIRUS; TOBACCO; TRANSGENES; VIRAL NONSTRUCTURAL PROTEINS;

EID: 33749048589     PISSN: 0001723X     EISSN: None     Source Type: Journal    
DOI: None     Document Type: Article

References (11)

1. Bubner B, Gase K, Baldwin IT (2004): Two-fold differences are the detection limit for determining transgene copy numbers in plants by real-time PCR. BMC Biotechnology 4, 14 (http://www.biomedcentral.eom/1472- 6750/4/14).
2. Hobbs SLA, Warkentin TD, DeLong CMO (1993): Transgene copy number can be positively or negatively associated with transgene expression. Plant Mol. Biol. 21, 17-26.
3. Ingham DJ, Beer S, Money S, Hansen G (2001): Quantitative Real-Time PCR Assay for Determining Transgene Copy Number in Transformed Plants. BioTechniques 31, 132-140.
4. Li Z, Hansen JL, Liu Y, Zemetra RS, Berger PH (2004): Using Real-Time PCR to Determine Transgene Copy Number in Wheat. Plant Mol. Biol. Rep. 22, 179-188.
5. -ΔΔCt method. Methods 25, 402-408.
6. Mason G, Provero P, Vaira AM, Accotto GP (2002): Estimating the number of integrations in transformed plants by quantitative real-time PCR. BMC Biotechnology 2, 20 (http://www.biomedcentral.com/1472-6750/2/20).
7. Nováková S, Mazúrová L', Čeř ovská N, Šubr ZW (2005): Transgenic tobacco plants carrying the non-structural P3 gene of potato virus A. Biol. Plant. 49, 593-598.
8. Puurand U, Makinen K, Paulin L, Saarma M (1994): The nucleotide sequence of potato virus A genomic RNA and its sequence similarities with other potyviruses. J. Gen. Virol. 75, 457-461
9. Sambrook J, Fritsch EF, Maniatis T (1989): Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, New York.
10. Topping JF, Lindsey K (1997): Molecular characterization of transformed plants. In Clark MS (Ed.): Plant Molecular Biology. A Laboratory Manual. Springer-Verlag, Berlin-Heidelberg-New York, pp. 427442.
11. Waiden R, Hayashi H, Lubenow H, Czaja I, Schell J (1994): Auxin inducibility and developmental expression of axi1: a gene directing auxin independent growth in tobacco protoplasts. EMBO J. 13, 4729-4736.