The molecular characterization of a gamma-ray induced mutation resulting in altered HLA-B5 gene expression
Thesis/Dissertation
·
OSTI ID:7254582
The preliminary analysis of LCL 721.53 revealed the presence of the HLA-class I B5 gene at the DNA level, the marked reduction of HLA-B5 expression at the mRNA level, and the absence of the HLA-B5 product at the protein level. The HLA-B5 gene of 721.53 was thus chosen as a model for the molecular characterization of the effects of gamma-irradiation on HLA-class I gene expression. The 6.8 kb EcoRI fragment containing the HLA-B5 gene of 721.53 was cloned into {lambda}gt11. Hybrids of the HLA-B5 gene cloned from 721.53 and 721 were constructed, transfected into mouse L cells, and assayed for the presence of HLA-B5 mRNA. The mutation resulting in the absence of steady-state HLA-B5 mRNA in 721.53 lay within the first 1.6 kb of the gene. Sanger dideoxy sequencing of the 1.6 kb EcoRI-KpnI fragment of both 721.53 and 721 revealed a deletion of 22 bp in the {alpha}1 coding region, encompassing amino acids 76 through 83. This frameshift mutation in 721.53 results in the formation of three in frame stop codons in the {alpha}2 exon.
- Research Organization:
- Minnesota Univ., Minneapolis, MN (USA)
- OSTI ID:
- 7254582
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
560120* -- Radiation Effects on Biochemicals
Cells
& Tissue Culture
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
AMINO ACID SEQUENCE
ANIMALS
ELECTROMAGNETIC RADIATION
GAMMA RADIATION
GENE MUTATIONS
GENE REGULATION
GENES
IONIZING RADIATIONS
MAMMALS
MESSENGER-RNA
MICE
MOLECULAR STRUCTURE
MUTATIONS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
RADIATIONS
RADIOINDUCTION
RNA
RODENTS
TRANSCRIPTION
VERTEBRATES
Cells
& Tissue Culture
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
AMINO ACID SEQUENCE
ANIMALS
ELECTROMAGNETIC RADIATION
GAMMA RADIATION
GENE MUTATIONS
GENE REGULATION
GENES
IONIZING RADIATIONS
MAMMALS
MESSENGER-RNA
MICE
MOLECULAR STRUCTURE
MUTATIONS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
RADIATIONS
RADIOINDUCTION
RNA
RODENTS
TRANSCRIPTION
VERTEBRATES