title: Beta-thalassemia (beta 90 GAG-->TAG): genetic diagnosis by allele-specific polymerase chain reaction and estimation of mutant mRNA expression by reverse transcription polymerase chain reaction.
abstract: Genetic diagnosis of a family of beta-thalassemia (beta 90 GAG-->TAG) was carried out by allele-specific polymerase chain reaction (AS-PCR). The proband, her daughter and granddaughter were proved to be heterozygotes of normal and mutant alleles. As some nonsense mutations express a decreased amount of MrNAs, we determined the expression level of the mutant mRNA of beta-thalassemia (beta 90 GAG-->TAG), by application of a combination method of reverse transcription PCR (RT-PCR) and dot-blot hybridization with allele-specific oligonucleotides. The mutant mRNA was not markedly reduced. In conclusion, 1) individuals with the mutant beta-globin gene were diagnosed successfully by AS-PCR, and 2) a significant amount of the mutant beta-globin mRNA was synthesized.
authors.affiliation: First Department of Medicine, Wakayama University of Medical Science.
authors.forename: K
authors.initials: K
authors.lastname: Ekawa
authors.forename: M
authors.initials: M
authors.lastname: Nishi
authors.forename: T
authors.initials: T
authors.lastname: Okamura
authors.forename: Y
authors.initials: Y
authors.lastname: Nomura
authors.forename: T
authors.initials: T
authors.lastname: Sanke
authors.forename: K
authors.initials: K
authors.lastname: Nanjo
mesh_terms: D000483:Alleles; D005260:Female; D015870:Gene Expression; D006580:Genetic Carrier Screening; D005914:Globins / Q000235:genetics*; D006801:Humans; D008297:Male; D008875:Middle Aged; D009693:Nucleic Acid Hybridization; D010375:Pedigree; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000096:biosynthesis*; D012194:RNA-Directed DNA Polymerase; D017086:beta-Thalassemia / Q000175:diagnosis / Q000235:genetics*
chemical_list: D012333:RNA, Messenger; D005914:Globins; D012194:RNA-Directed DNA Polymerase