RESUMO
OBJECTIVE: To explore the molecular mechanism involved in patient with congenital FV deficiency. METHODS: Activity of FV was determined by biochemical method. The PCR products of FV gene was analyzed by DNA sequencing directly or cloned into T-vector prior to DNA analysis. The mutation of FV gene in proband and his family numbers was analysed by restriction enzyme analysis. Its occurrence was investigated in the control group. DNA was extracted from the peripheral blood mono1nuclear cells of the proband, male, 18 years old, and his parents. The PCR products were analyzed by direct sequencing or cloned into T-vector prior to DNA analysis. One hundred patients with different kind of hemotopathy were used as controls. RESULTS: A single point mutation, AG-->GG was found at position 3' splice site of intron 8 of the proband. This mutation was confirmed by family screening. CONCLUSION: A single point mutation, AG-->GG at position 3' splice site of intron 8 mutation of FV gene is related to the pathogenesis of congenital FV deficiency.
Assuntos
Coagulação Sanguínea/genética , Deficiência do Fator V/genética , Fator V/genética , Sítios de Splice de RNA , Splicing de RNA/genética , Adolescente , Análise Mutacional de DNA , Deficiência do Fator V/fisiopatologia , Feminino , Humanos , Íntrons , Masculino , Pessoa de Meia-Idade , Mutação PuntualRESUMO
OBJECTIVE: To explore the molecular mechanisms involved in the patient with congenital FV deficiency. METHODS: Activity of FV was determined by biochemical method. The PCR products of FV gene was analysed by directly sequencing or sequencing after cloned into T-vector. The mutative FV gene was analysed by restriction enzyme analysis in the proband and her family members. RESULTS: A homozygous missense mutation G5729T resulting in Gly1880Val was revealed in the proband and confirmed in the family screening. Structure-function studies of the factor V mutants (Gly1880Val) demonstrated the importance of Gly1880 for structural stability of the Factor V. CONCLUSION: G5729T mutation of FV gene is related to the pathogenesis of congenital FV deficiency.