Mapping of class I DNA sequences within the human major histocompatibility complex

Harry T. Orr, Robert DeMars

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Mutants that had lost expression of alleles of one or more HLA loci were isolated with immunoselection after γ-irradiation of a human lymphoblastoid cell line LCL 721. DNAs from the mutants were digested with restriction endonucleases and analyzed by Southern blotting using probes for class I HLA genes. Eight polymorphic cut sites for HindIII and PvuII were discovered in class I-associated sequences of LCL 721. Losses of specific fragments generated by restriction enzymes could be associated with losses of specific antigenic expressions and it was possible in this way to assign HLA-Al, HLA-A2, and HLA-B8 to specific DNA fragments. Patterns of γ-ray-induced segregations of DNA fragments permitted rough linkage alignment of about 30% of the fragments generated by PvuII. The resultant map showed that there are class I HLA genes on the telomeric side of the HLA-A locus. Restriction enzyme site polymorphisms were also examined in a panel of DNAs isolated from peripheral blood lymphocytes (PBLs) of HLA-typed individuals. This panel of PBL DNA complemented the analysis using the HLA deletion mutants.

Original languageEnglish (US)
Pages (from-to)489-502
Number of pages14
JournalImmunogenetics
Volume18
Issue number5
DOIs
StatePublished - Nov 1983

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Major Histocompatibility Complex
DNA
MHC Class I Genes
HLA-B8 Antigen
Lymphocytes
HLA-A2 Antigen
HLA-A Antigens
DNA Restriction Enzymes
Enzymes
Southern Blotting
Alleles
Cell Line

Cite this

Mapping of class I DNA sequences within the human major histocompatibility complex. / Orr, Harry T.; DeMars, Robert.

In: Immunogenetics, Vol. 18, No. 5, 11.1983, p. 489-502.

Research output: Contribution to journalArticle

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