scholarly journals Use of oligonucleotide hybridization in the characterization of a beta zero-thalassemia gene (beta 37 TGG----TGA) in a Saudi Arabian family [published erratum appears in Blood 1986 Jul;68(1):323]

Blood ◽  
1986 ◽  
Vol 67 (4) ◽  
pp. 1185-1188
Author(s):  
CD Boehm ◽  
CE Dowling ◽  
PG Waber ◽  
PJ Giardina ◽  
HH Jr Kazazian
Keyword(s):  
Genomic Dna ◽  
Globin Gene ◽  
Point Mutations ◽  
Saudi Arabian ◽  
Globin Genes ◽  
Beta Globin ◽  
Nucleotide Substitutions ◽  
Exon 2 ◽  
Oligonucleotide Hybridization

Analysis of restriction site polymorphisms in the beta-globin gene cluster of a Saudi Arabian female with beta zero-thalassemia demonstrated that both of her beta-globin genes were missing a nonpolymorphic AvaII site in exon 2. Examination of the normal nucleotide sequence surrounding this AvaII site revealed that either of two nucleotide substitutions, TGG----TAG or TGG----TGA, could produce a nonsense codon at codon 37 and eliminate the AvaII site. Consequently, two oligonucleotides (19-mers spanning codons 36 through 41 and containing either TAG or TGA at codon 37) were synthesized and hybridized against genomic DNA of the proband and her family. Specific hybridization with one of the oligomers demonstrated that the patient's beta o-thalassemia was the result of homozygosity for the TGG----TGA mutation at codon 37. In certain cases, oligonucleotide hybridization using genomic DNA may obviate the need for gene cloning and sequencing in the characterization of point mutations.

scholarly journals Use of oligonucleotide hybridization in the characterization of a beta zero-thalassemia gene (beta 37 TGG----TGA) in a Saudi Arabian family [published erratum appears in Blood 1986 Jul;68(1):323]

Blood ◽  
1986 ◽  
Vol 67 (4) ◽  
pp. 1185-1188 ◽  
Author(s):  
CD Boehm ◽  
CE Dowling ◽  
PG Waber ◽  
PJ Giardina ◽  
HH Jr Kazazian
Keyword(s):  
Genomic Dna ◽  
Globin Gene ◽  
Point Mutations ◽  
Saudi Arabian ◽  
Globin Genes ◽  
Beta Globin ◽  
Nucleotide Substitutions ◽  
Exon 2 ◽  
Oligonucleotide Hybridization

Abstract Analysis of restriction site polymorphisms in the beta-globin gene cluster of a Saudi Arabian female with beta zero-thalassemia demonstrated that both of her beta-globin genes were missing a nonpolymorphic AvaII site in exon 2. Examination of the normal nucleotide sequence surrounding this AvaII site revealed that either of two nucleotide substitutions, TGG----TAG or TGG----TGA, could produce a nonsense codon at codon 37 and eliminate the AvaII site. Consequently, two oligonucleotides (19-mers spanning codons 36 through 41 and containing either TAG or TGA at codon 37) were synthesized and hybridized against genomic DNA of the proband and her family. Specific hybridization with one of the oligomers demonstrated that the patient's beta o-thalassemia was the result of homozygosity for the TGG----TGA mutation at codon 37. In certain cases, oligonucleotide hybridization using genomic DNA may obviate the need for gene cloning and sequencing in the characterization of point mutations.

scholarly journals Clinical and genetic heterogeneity in black patients with homozygous beta-thalassemia from the southeastern United States

Blood ◽  
1988 ◽  
Vol 72 (3) ◽  
pp. 1007-1014 ◽  
Author(s):  
JM Gonzalez-Redondo ◽  
TA Stoming ◽  
KD Lanclos ◽  
YC Gu ◽  
A Kutlar ◽  
...  
Keyword(s):  
South Carolina ◽  
Genomic Dna ◽  
Stop Codon ◽  
Globin Gene ◽  
Beta Thalassemia ◽  
Beta Globin ◽  
Nucleotide Substitutions ◽  
Taq Polymerase ◽  
Black Patients ◽  
Codon 61

Abstract The presence of various substitutions and deletions resulting in beta- thalassemia was studied in 19 black patients with homozygous beta- thalassemia and in numerous relatives; all patients were from Georgia, South Carolina, and Alabama. Methodology included gene mapping, amplification of genomic DNA with Taq polymerase, identification of known nucleotide substitutions or a single nucleotide deletion through hybridization with synthetic oligonucleotides, cloning and sequencing of a beta-globin gene, and sequencing of amplified genomic DNA. Of the 38 chromosomes tested, 21 (55%) had the A----G substitution at nt -29, eight (21%) had the C----T substitution at nt -88, three (8%) had the substitution at codon 24, while one each of the following abnormalities were also detected: frameshift at codon 6, a C----A mutation at nt 848 of the beta IVS-II (new), an A----T mutation at codon 61 (new), a deletion of 1.35 kilobases including the 5′ end of beta, a Ggamma(Agammadelta beta) degree-thalassemia, and one thalassemia determinant that remained unidentified. The C----A mutation at nt 848 of IVS-II occurred at a position 3 nucleotides 5′ to the third exon, adjacent to the invariant AG dinucleotide of the acceptor sequence. The A----T mutation in codon 61 (AAG----TAG) resulted in the creation of a stop codon and thus in beta degree-thalassemia. The various mutations occurred on chromosomes with different haplotypes; however, chromosomes with a specific mutation but with different haplotypes belonged to one specific framework, which suggested that crossovers were responsible for these different types. Hemoglobin (Hb) F levels were generally high (55% to 75% with 98.5% in one patient with beta degree/beta degree); a few patients with specific haplotypes and an alpha-thalassemia-2 heterozygosity had a lower Hb F level. The Ggamma in the Hb F was consistently high when the C----T mutation occurred at nt -158 to the Cap site of the Ggamma-globin gene; seven patients with +/+ at this site had an average Ggamma of 73.8%, eight patients with +/- had 64.8%, and one patient with -/- had 34.2%. Variations in hematologic values and in Hb F, Ggamma, and Hb A2 levels of relatives with a beta- thalassemia heterozygosity depended to some extent on the types of mutations or deletions and on the haplotypes of the chromosomes with the beta-thalassemia determinant.

scholarly journals SHORT GENE CONVERSIONS IN THE HUMAN FETAL GLOBIN GENE REGION: A BY-PRODUCT OF CHROMOSOME PAIRING DURING MEIOSIS?

Genetics ◽  
1986 ◽  
Vol 112 (2) ◽  
pp. 343-358
Author(s):  
Patricia A Powers ◽  
Oliver Smithies
Keyword(s):  
Chromosome Pairing ◽  
Dna Sequences ◽  
Meiotic Chromosome ◽  
Globin Gene ◽  
Point Mutations ◽  
Globin Genes ◽  
Nucleotide Substitutions ◽  
Sequence Comparisons ◽  
The One ◽  
Gene Conversions

ABSTRACT DNA sequence comparisons of a 1200-base pair (bp) region in 14 human fetal globin genes in seven linked pairs reveal 31 nucleotide substitutions at positions where the fetal globin genes, G  y and A  y, usually differ. In each case, the newly substituted nucleotide is identical to the one found at the same position in the linked nonallelic gene. Most of these nucleotide substitutions are clearly the result of gene conversions, but 11 could be the result of either very short gene conversions or of point mutations. The unexpectedly frequent occurrence of these short gene conversions suggests that they may be the relics of some normal interaction between homologous but nonallelic DNA sequences, and we discuss the possibility that they result from interactions occurring between homologous sequences during the process of meiotic chromosome pairing.

scholarly journals Clinical and genetic heterogeneity in black patients with homozygous beta-thalassemia from the southeastern United States

Blood ◽  
1988 ◽  
Vol 72 (3) ◽  
pp. 1007-1014 ◽  
Author(s):  
JM Gonzalez-Redondo ◽  
TA Stoming ◽  
KD Lanclos ◽  
YC Gu ◽  
A Kutlar ◽  
...  
Keyword(s):  
South Carolina ◽  
Genomic Dna ◽  
Stop Codon ◽  
Globin Gene ◽  
Beta Thalassemia ◽  
Beta Globin ◽  
Nucleotide Substitutions ◽  
Taq Polymerase ◽  
Black Patients ◽  
Codon 61

The presence of various substitutions and deletions resulting in beta- thalassemia was studied in 19 black patients with homozygous beta- thalassemia and in numerous relatives; all patients were from Georgia, South Carolina, and Alabama. Methodology included gene mapping, amplification of genomic DNA with Taq polymerase, identification of known nucleotide substitutions or a single nucleotide deletion through hybridization with synthetic oligonucleotides, cloning and sequencing of a beta-globin gene, and sequencing of amplified genomic DNA. Of the 38 chromosomes tested, 21 (55%) had the A----G substitution at nt -29, eight (21%) had the C----T substitution at nt -88, three (8%) had the substitution at codon 24, while one each of the following abnormalities were also detected: frameshift at codon 6, a C----A mutation at nt 848 of the beta IVS-II (new), an A----T mutation at codon 61 (new), a deletion of 1.35 kilobases including the 5′ end of beta, a Ggamma(Agammadelta beta) degree-thalassemia, and one thalassemia determinant that remained unidentified. The C----A mutation at nt 848 of IVS-II occurred at a position 3 nucleotides 5′ to the third exon, adjacent to the invariant AG dinucleotide of the acceptor sequence. The A----T mutation in codon 61 (AAG----TAG) resulted in the creation of a stop codon and thus in beta degree-thalassemia. The various mutations occurred on chromosomes with different haplotypes; however, chromosomes with a specific mutation but with different haplotypes belonged to one specific framework, which suggested that crossovers were responsible for these different types. Hemoglobin (Hb) F levels were generally high (55% to 75% with 98.5% in one patient with beta degree/beta degree); a few patients with specific haplotypes and an alpha-thalassemia-2 heterozygosity had a lower Hb F level. The Ggamma in the Hb F was consistently high when the C----T mutation occurred at nt -158 to the Cap site of the Ggamma-globin gene; seven patients with +/+ at this site had an average Ggamma of 73.8%, eight patients with +/- had 64.8%, and one patient with -/- had 34.2%. Variations in hematologic values and in Hb F, Ggamma, and Hb A2 levels of relatives with a beta- thalassemia heterozygosity depended to some extent on the types of mutations or deletions and on the haplotypes of the chromosomes with the beta-thalassemia determinant.

scholarly journals Beta-thalassemia due to two novel nucleotide substitutions in consensus acceptor splice sequences of the beta-globin gene

Blood ◽  
1989 ◽  
Vol 73 (4) ◽  
pp. 914-918
Author(s):  
C Wong ◽  
SE Antonarakis ◽  
SC Goff ◽  
SH Orkin ◽  
BG Forget ◽  
...  
Keyword(s):  
Splice Site ◽  
Rna Splicing ◽  
Globin Gene ◽  
Beta Thalassemia ◽  
Beta Globin ◽  
Nucleotide Substitutions ◽  
Splice Site Selection ◽  
Exon 2 ◽  
Consensus Sequences ◽  
Beta Globin Gene

We have identified two novel RNA-splicing mutations affecting a critical nucleotide (nt) in the acceptor consensus sequences at both the IVS-1/exon 2 and IVS-2/exon 3 junctions of the human beta-globin gene. Both mutations are single nt substitutions, T to G and C to A, at position -3 adjacent to the invariant AG dinucleotide. For the IVS- 2/exon 3 mutation abnormal splicing into the cryptic splice site at IVS- 2 nt 579 is documented. Identification of these two mutations provides further support for the importance of the location of specific nucleotides within the consensus sequences in splice site selection and RNA processing.

scholarly journals Molecular and hematologic characterization of Scottish-Irish type (epsilon gamma delta beta)zero thalassemia

Blood ◽  
1990 ◽  
Vol 76 (10) ◽  
pp. 2132-2138
Author(s):  
RJ Trent ◽  
BG Williams ◽  
A Kearney ◽  
T Wilkinson ◽  
PC Harris
Keyword(s):  
Dna Sequence ◽  
Globin Gene ◽  
Globin Genes ◽  
The Novel ◽  
Beta Globin ◽  
Gamma Delta ◽  
Beta Globin Gene ◽  
Clinical Phenotypes ◽  
Dna Deletion

The DNA deletion associated with an example of (epsilon gamma delta beta)zero thalassemia (Scottish-Irish type) was characterized. The deletion is approximately 205 kb in length and involves the epsilon, G gamma, A gamma, delta, and beta globin genes. The breakpoint is located 263 bp 3′ to exon 3 of the beta globin gene. An LI (KpnI) repeat element approximately 320 bp in size is found at the 3′ end of the novel DNA sequence. Different clinical phenotypes for three heterozygous neonates suggest that the deletion alone does not predict severity of (epsilon gamma delta beta)zero thalassemia at this age.

scholarly journals Beta-thalassemia due to two novel nucleotide substitutions in consensus acceptor splice sequences of the beta-globin gene

Blood ◽  
1989 ◽  
Vol 73 (4) ◽  
pp. 914-918 ◽  
Author(s):  
C Wong ◽  
SE Antonarakis ◽  
SC Goff ◽  
SH Orkin ◽  
BG Forget ◽  
...  
Keyword(s):  
Splice Site ◽  
Rna Splicing ◽  
Globin Gene ◽  
Beta Thalassemia ◽  
Beta Globin ◽  
Nucleotide Substitutions ◽  
Splice Site Selection ◽  
Exon 2 ◽  
Consensus Sequences ◽  
Beta Globin Gene

Abstract We have identified two novel RNA-splicing mutations affecting a critical nucleotide (nt) in the acceptor consensus sequences at both the IVS-1/exon 2 and IVS-2/exon 3 junctions of the human beta-globin gene. Both mutations are single nt substitutions, T to G and C to A, at position -3 adjacent to the invariant AG dinucleotide. For the IVS- 2/exon 3 mutation abnormal splicing into the cryptic splice site at IVS- 2 nt 579 is documented. Identification of these two mutations provides further support for the importance of the location of specific nucleotides within the consensus sequences in splice site selection and RNA processing.

scholarly journals A deletion/inversion rearrangement of the beta-globin gene cluster in a Turkish family with delta beta zero-thalassemia intermedia

Blood ◽  
1992 ◽  
Vol 79 (9) ◽  
pp. 2455-2459
Author(s):  
AE Kulozik ◽  
A Bellan-Koch ◽  
E Kohne ◽  
E Kleihauer
Keyword(s):  
Gene Cluster ◽  
Globin Gene ◽  
Point Mutations ◽  
Gene Promoters ◽  
Globin Genes ◽  
Beta Globin ◽  
Beta Globin Gene ◽  
Gamma Globin ◽  
Globin Gene Cluster

The most common forms of hereditary persistence of fetal hemoglobin synthesis (HPFH) and delta beta zero-thalassemia result from simple deletions of the beta-globin gene cluster or from point mutations in the gamma-globin gene promoters. These naturally occurring mutants extend our understanding of globin gene regulation and hemoglobin switching. Furthermore, they provide the opportunity to test in vivo hypothetical switching models that are based on the experimental approach. We report here a family with delta beta zero-thalassemia from Turkey with a complex rearrangement of the beta-globin gene cluster that involves two deletions of 11.5 kb and 1.6 kb, and an inversion of 7.6 kb. The larger deletion removes both the delta-and the beta-globin genes with 3′ flanking sequences, whereas the smaller deletion affects DNA of unknown function. The inversion contains the entire L1 repeat 3′ of the beta-globin gene. There are structural motifs near the breakpoints (introduction of an “orphan” nucleotide, multiple direct and inverted repeats) suggesting a nonhomologous type of recombination event. The hematologic phenotype and the molecular structure of the rearranged beta-globin gene cluster are consistent with a competitive relationship between the fetal and the adult globin genes and/or with the translocation of enhancer sequences into the gamma-globin gene region.

scholarly journals Molecular and hematologic characterization of Scottish-Irish type (epsilon gamma delta beta)zero thalassemia

Blood ◽  
1990 ◽  
Vol 76 (10) ◽  
pp. 2132-2138 ◽  
Author(s):  
RJ Trent ◽  
BG Williams ◽  
A Kearney ◽  
T Wilkinson ◽  
PC Harris
Keyword(s):  
Dna Sequence ◽  
Globin Gene ◽  
Globin Genes ◽  
The Novel ◽  
Beta Globin ◽  
Gamma Delta ◽  
Beta Globin Gene ◽  
Clinical Phenotypes ◽  
Dna Deletion

Abstract The DNA deletion associated with an example of (epsilon gamma delta beta)zero thalassemia (Scottish-Irish type) was characterized. The deletion is approximately 205 kb in length and involves the epsilon, G gamma, A gamma, delta, and beta globin genes. The breakpoint is located 263 bp 3′ to exon 3 of the beta globin gene. An LI (KpnI) repeat element approximately 320 bp in size is found at the 3′ end of the novel DNA sequence. Different clinical phenotypes for three heterozygous neonates suggest that the deletion alone does not predict severity of (epsilon gamma delta beta)zero thalassemia at this age.

scholarly journals A deletion/inversion rearrangement of the beta-globin gene cluster in a Turkish family with delta beta zero-thalassemia intermedia

Blood ◽  
1992 ◽  
Vol 79 (9) ◽  
pp. 2455-2459 ◽  
Author(s):  
AE Kulozik ◽  
A Bellan-Koch ◽  
E Kohne ◽  
E Kleihauer
Keyword(s):  
Gene Cluster ◽  
Globin Gene ◽  
Point Mutations ◽  
Gene Promoters ◽  
Globin Genes ◽  
Beta Globin ◽  
Beta Globin Gene ◽  
Gamma Globin ◽  
Globin Gene Cluster

Abstract The most common forms of hereditary persistence of fetal hemoglobin synthesis (HPFH) and delta beta zero-thalassemia result from simple deletions of the beta-globin gene cluster or from point mutations in the gamma-globin gene promoters. These naturally occurring mutants extend our understanding of globin gene regulation and hemoglobin switching. Furthermore, they provide the opportunity to test in vivo hypothetical switching models that are based on the experimental approach. We report here a family with delta beta zero-thalassemia from Turkey with a complex rearrangement of the beta-globin gene cluster that involves two deletions of 11.5 kb and 1.6 kb, and an inversion of 7.6 kb. The larger deletion removes both the delta-and the beta-globin genes with 3′ flanking sequences, whereas the smaller deletion affects DNA of unknown function. The inversion contains the entire L1 repeat 3′ of the beta-globin gene. There are structural motifs near the breakpoints (introduction of an “orphan” nucleotide, multiple direct and inverted repeats) suggesting a nonhomologous type of recombination event. The hematologic phenotype and the molecular structure of the rearranged beta-globin gene cluster are consistent with a competitive relationship between the fetal and the adult globin genes and/or with the translocation of enhancer sequences into the gamma-globin gene region.

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