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Modulation of factor VII levels by intron 7 polymorphisms: population and in vitro studies by Mirko Pinotti, Raffaella Toso, Domenico Girelli, Debora Bindini,

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Presentation on theme: "Modulation of factor VII levels by intron 7 polymorphisms: population and in vitro studies by Mirko Pinotti, Raffaella Toso, Domenico Girelli, Debora Bindini,"— Presentation transcript:

1 Modulation of factor VII levels by intron 7 polymorphisms: population and in vitro studies
by Mirko Pinotti, Raffaella Toso, Domenico Girelli, Debora Bindini, Paolo Ferraresi, Maria L. Papa, Roberto Corrocher, Giovanna Marchetti, and Francesco Bernardi Blood Volume 95(11): June 1, 2000 ©2000 by American Society of Hematology

2 Mirko Pinotti et al. Blood 2000;95:3423-3428
©2000 by American Society of Hematology

3 Mirko Pinotti et al. Blood 2000;95:3423-3428
©2000 by American Society of Hematology

4 Schematic diagram of the F7 gene and mRNA regions expressed in mammalian cells.(A) pcDNA3 plasmid (pIVS7H5, pIVS7H6, pIVS7H7, pIVS7H8, pIVS7 + 5A, and pIVS7 + 7G) containing the different allelic forms ordered by repeat number. Schematic diagram of the F7 gene and mRNA regions expressed in mammalian cells.(A) pcDNA3 plasmid (pIVS7H5, pIVS7H6, pIVS7H7, pIVS7H8, pIVS7 + 5A, and pIVS7 + 7G) containing the different allelic forms ordered by repeat number. (B) Spliced transcription products from plasmids and the size of RT-PCR–amplified fragments. pIVS7 + 5A, used as an internal control, produces the altered transcript (+5A), which contains an additional 37-bp sequence (indicated by filled square). Mirko Pinotti et al. Blood 2000;95: ©2000 by American Society of Hematology

5 mRNA expression studies with RT-PCR–amplified fragments of FVII mRNA expressed in BHK cells cotransfected with pIVS7H5 and pIVS7 + 5A (internal control).The following pIVS7 + 5A/pIVS7H5 DNA concentration ratios were used in lanes 1-4: 1:1, 1:0.3, 1:0.15, an... mRNA expression studies with RT-PCR–amplified fragments of FVII mRNA expressed in BHK cells cotransfected with pIVS7H5 and pIVS7 + 5A (internal control).The following pIVS7 + 5A/pIVS7H5 DNA concentration ratios were used in lanes 1-4: 1:1, 1:0.3, 1:0.15, and 1:0.1, respectively. Mirko Pinotti et al. Blood 2000;95: ©2000 by American Society of Hematology

6 Relative expression at the mRNA level of the differentIVS7 allelic forms.Cotransfections with pIVS7 + 5A and investigated constructs at concentration ratios of 1:0.1 (black columns) and 1:0.15 (gray columns) were carried out. Relative expression at the mRNA level of the differentIVS7 allelic forms.Cotransfections with pIVS7 + 5A and investigated constructs at concentration ratios of 1:0.1 (black columns) and 1:0.15 (gray columns) were carried out. Mirko Pinotti et al. Blood 2000;95: ©2000 by American Society of Hematology

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