The molecular characterization of a gamma-ray induced mutation resulting in altered HLA-B5 gene expression

Miscellaneous · Fri Jan 01 00:00:00 EST 1988

OSTI ID:7254582

Bowers, A C

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
• Resource Relation: Other Information: Thesis (Ph. D.)
• Country of Publication: United States
• Language: English

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