BIOMAT Database Logo BIOMAT Database Logo

Rapid identification by denaturing gradient gel electrophoresis of mutations in the gamma-globin gene promoters in non-deletion type HPFH. 1992

E Gottardi, and M Losekoot, and R Fodde, and G Saglio, and C Camaschella, and L F Bernini
Dipartimento di Scienze Biomediche e Oncologia Umana, Università di Torino, Italy.

We have outlined a fast, non-radioactive strategy to identify point mutations in the 5' flanking region of the gamma-globin genes using denaturing gradient gel electrophoresis (DGGE) of amplified DNA. In a sample of previously characterized carriers of non deletion-type hereditary persistence of fetal haemoglobin (HPFH) the different point mutations in both gamma gene promoters could be easily identified by DGGE of a 327 bp fragment. A 4 bp deletion at position -225 to -222 of the A gamma globin gene was unexpectedly found in several samples and shown to represent a frequent polymorphism. Analysis of a 681 bp fragment specific for the 5' region of the A gamma gene, showed that this can be used to determine the haplotype of the chromosome under study. This technique may be useful in the study of sequence variations associated with high Hb F expression in physiological and pathological conditions.

UI MeSH Term Description Entries
D009154 Mutation Any detectable and heritable change in the genetic material that causes a change in the GENOTYPE and which is transmitted to daughter cells and to succeeding generations. Mutations
D011401 Promoter Regions, Genetic DNA sequences which are recognized (directly or indirectly) and bound by a DNA-dependent RNA polymerase during the initiation of transcription. Highly conserved sequences within the promoter include the Pribnow box in bacteria and the TATA BOX in eukaryotes. rRNA Promoter,Early Promoters, Genetic,Late Promoters, Genetic,Middle Promoters, Genetic,Promoter Regions,Promoter, Genetic,Promotor Regions,Promotor, Genetic,Pseudopromoter, Genetic,Early Promoter, Genetic,Genetic Late Promoter,Genetic Middle Promoters,Genetic Promoter,Genetic Promoter Region,Genetic Promoter Regions,Genetic Promoters,Genetic Promotor,Genetic Promotors,Genetic Pseudopromoter,Genetic Pseudopromoters,Late Promoter, Genetic,Middle Promoter, Genetic,Promoter Region,Promoter Region, Genetic,Promoter, Genetic Early,Promoter, rRNA,Promoters, Genetic,Promoters, Genetic Middle,Promoters, rRNA,Promotor Region,Promotors, Genetic,Pseudopromoters, Genetic,Region, Genetic Promoter,Region, Promoter,Region, Promotor,Regions, Genetic Promoter,Regions, Promoter,Regions, Promotor,rRNA Promoters
D011489 Protein Denaturation Disruption of the non-covalent bonds and/or disulfide bonds responsible for maintaining the three-dimensional shape and activity of the native protein. Denaturation, Protein,Denaturations, Protein,Protein Denaturations
D004252 DNA Mutational Analysis Biochemical identification of mutational changes in a nucleotide sequence. Mutational Analysis, DNA,Analysis, DNA Mutational,Analyses, DNA Mutational,DNA Mutational Analyses,Mutational Analyses, DNA
D004591 Electrophoresis, Polyacrylamide Gel Electrophoresis in which a polyacrylamide gel is used as the diffusion medium. Polyacrylamide Gel Electrophoresis,SDS-PAGE,Sodium Dodecyl Sulfate-PAGE,Gel Electrophoresis, Polyacrylamide,SDS PAGE,Sodium Dodecyl Sulfate PAGE,Sodium Dodecyl Sulfate-PAGEs
D005319 Fetal Hemoglobin The major component of hemoglobin in the fetus. This HEMOGLOBIN has two alpha and two gamma polypeptide subunits in comparison to normal adult hemoglobin, which has two alpha and two beta polypeptide subunits. Fetal hemoglobin concentrations can be elevated (usually above 0.5%) in children and adults affected by LEUKEMIA and several types of ANEMIA. Hemoglobin F,Hemoglobin, Fetal
D005914 Globins A superfamily of proteins containing the globin fold which is composed of 6-8 alpha helices arranged in a characterstic HEME enclosing structure. Globin
D006453 Hemoglobinopathies A group of inherited disorders characterized by structural alterations within the hemoglobin molecule. Hemoglobinopathy
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D013789 Thalassemia A group of hereditary hemolytic anemias in which there is decreased synthesis of one or more hemoglobin polypeptide chains. There are several genetic types with clinical pictures ranging from barely detectable hematologic abnormality to severe and fatal anemia. Thalassemias

Related Publications

E Gottardi, and M Losekoot, and R Fodde, and G Saglio, and C Camaschella, and L F Bernini
Detecting prion protein gene mutations by denaturing gradient gel electrophoresis.
January 1994, Human mutation,
E Gottardi, and M Losekoot, and R Fodde, and G Saglio, and C Camaschella, and L F Bernini
Rapid detection of the highly polymorphic beta globin framework by denaturing gradient gel electrophoresis.
August 1992, Journal of medical genetics,
E Gottardi, and M Losekoot, and R Fodde, and G Saglio, and C Camaschella, and L F Bernini
Detection of mutations by denaturing gradient gel electrophoresis.
May 2001, Current protocols in human genetics,
E Gottardi, and M Losekoot, and R Fodde, and G Saglio, and C Camaschella, and L F Bernini
Identification of the multiple beta-thalassemia mutations by denaturing gradient gel electrophoresis.
February 1990, The Journal of clinical investigation,
E Gottardi, and M Losekoot, and R Fodde, and G Saglio, and C Camaschella, and L F Bernini
Detection of mutations in insulin receptor gene by denaturing gradient gel electrophoresis.
April 1992, Diabetes,
E Gottardi, and M Losekoot, and R Fodde, and G Saglio, and C Camaschella, and L F Bernini
Comparative studies of nondeletional HPFH gamma-globin gene promoters.
July 1993, Experimental hematology,
E Gottardi, and M Losekoot, and R Fodde, and G Saglio, and C Camaschella, and L F Bernini
Characterization of four mutations in the neurofibromatosis type 1 gene by denaturing gradient gel electrophoresis (DGGE).
April 1994, Human molecular genetics,
E Gottardi, and M Losekoot, and R Fodde, and G Saglio, and C Camaschella, and L F Bernini
Rapid detection of beta-globin gene mutations and polymorphisms by temporal temperature gradient gel electrophoresis.
May 2003, Clinical chemistry,
E Gottardi, and M Losekoot, and R Fodde, and G Saglio, and C Camaschella, and L F Bernini
Identification of five novel inactivating mutations in the human thyroid peroxidase gene by denaturing gradient gel electrophoresis.
January 1995, Human mutation,
E Gottardi, and M Losekoot, and R Fodde, and G Saglio, and C Camaschella, and L F Bernini
Detection of p53 point mutations by double-gradient, denaturing gradient gel electrophoresis.
December 1997, Electrophoresis,
Copied contents to your clipboard!