Determination of neutral polymorphisms (frameworks) of the human beta globin gene in beta thalassaemias by PCR/DGGE

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dc.contributor.authorMoreira, Haroldo Wilson [UNESP]
dc.contributor.authorDe Oliveira, Cristina Marta [UNESP]
dc.contributor.authorMartins, Carmen Silvia Bertuzzo
dc.contributor.authorDe Sales, Teresa Sueko
dc.contributor.authorCosta, Fernando Ferreira
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.contributor.institutionUniversidade Estadual de Campinas (UNICAMP)
dc.date.accessioned2014-05-27T11:18:12Z
dc.date.available2014-05-27T11:18:12Z
dc.date.issued1997-02-01
dc.description.abstractPurpose: Considering the importance of type beta thalassaemias as hereditary syndromes of high significance in different populations of Mediterranean origin and, by extension, in the Brazilian population, the objective of the present study was to determine by PCR/DGGE the gene structures responsible for neutral polymorphisms (frameworks) observed in the human beta globin gene associated with the mutations responsible for type beta thalassaemias in a sample of the Brazilian population and, more specifically, of the population of the State of São Paulo. Patients and methods: Thirty individuals with beta thalassaemic mutations were analyzed: 22 mutations were in codon 39 (C->T), 5 in IVS1-110 (G->A), 2 in IVS1-6 (T->C) and 1 in IVS1-1 (G->A). DNA was extracted and selective amplification was performed by PCR extending from position IVS1 nt 46 to IVS2 nt 126 (474 pb). The product was then analyzed by polyacrylamide gel electrophoresis on a denaturing 10-60% urea/formamide gradient. Results: The results demonstrated that, as expected, the mutations responsible for type beta thalassaemia observed in this population are of Mediterranean origin, with 73% distribution represented by codon 39,17% by IVS1-110, 7% by IVS1-6 and 3% by IVS1-1. In turn, framework distribution seems to indicate a higher frequency of Fr 1-1 in codon 39 and IVS1-110, of Fr 1-3 in IVS1-6 and of Fr 1-2 in IVS1-1. Conclusions: These results permit us to conclude that gene amplification by PCR followed by DGGE is an appropriate method for the separation of DNA molecules that differ even by a single base change and therefore can be utilized to detect the alterations observed in the human beta globin gene. This methodology shows that, using only a pair of primers, it is possible to define the frameworks that are observed in the beta globin gene.en
dc.description.affiliationDepto. de Analises Clinicas/FCF UNESP
dc.description.affiliationNucleo Hematologie Hemoterapia da F. UNESP
dc.description.affiliationDepto. de Genet. Médica/FCM UNICAMP
dc.description.affiliationCtro. de Hematol. e Hemoterapia/FCM UNICAMP
dc.description.affiliation, P.O. Box 502, 14801-970 Araraquara S.P.
dc.description.affiliationDepto. de Analises Clinicas Faculdade Cie. Farmaceuticas do C. Univ. Estadual Paulista - UNESP, Rua Expedicionarios do Brasil, 1621, 14801-902 Araraquara Estado de S.P.
dc.description.affiliationUnespDepto. de Analises Clinicas/FCF UNESP
dc.description.affiliationUnespNucleo Hematologie Hemoterapia da F. UNESP
dc.description.affiliationUnespDepto. de Analises Clinicas Faculdade Cie. Farmaceuticas do C. Univ. Estadual Paulista - UNESP, Rua Expedicionarios do Brasil, 1621, 14801-902 Araraquara Estado de S.P.
dc.format.extent21-24
dc.identifier.citationSangre, v. 42, n. 1, p. 21-24, 1997.
dc.identifier.issn0036-4355
dc.identifier.scopus2-s2.0-0031063763
dc.identifier.urihttp://hdl.handle.net/11449/65039
dc.language.isoeng
dc.relation.ispartofSangre
dc.rights.accessRightsAcesso restrito
dc.sourceScopus
dc.subjectdithiothreitol
dc.subjectglobin
dc.subjectpapain
dc.subjectadolescent
dc.subjectadult
dc.subjectbeta thalassemia
dc.subjectBrazil
dc.subjectDNA denaturation
dc.subjectethnology
dc.subjectfemale
dc.subjectgenetic polymorphism
dc.subjectgenetics
dc.subjecthuman
dc.subjectItaly
dc.subjectmale
dc.subjectmiddle aged
dc.subjectpolyacrylamide gel electrophoresis
dc.subjectpolymerase chain reaction
dc.subjectPortugal
dc.subjectSpain
dc.subjectAdolescent
dc.subjectAdult
dc.subjectbeta-Thalassemia
dc.subjectDithiothreitol
dc.subjectElectrophoresis, Polyacrylamide Gel
dc.subjectFemale
dc.subjectGlobins
dc.subjectHumans
dc.subjectMale
dc.subjectMiddle Aged
dc.subjectNucleic Acid Denaturation
dc.subjectPapain
dc.subjectPolymerase Chain Reaction
dc.subjectPolymorphism, Genetic
dc.titleDetermination of neutral polymorphisms (frameworks) of the human beta globin gene in beta thalassaemias by PCR/DGGEen
dc.typeArtigo
unesp.campusUniversidade Estadual Paulista (Unesp), Faculdade de Ciências Farmacêuticas, Araraquarapt
unesp.departmentAnálises Clínicas - FCFpt

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Araraquara - FCF - Faculdade de Ciências Farmacêuticas