Isolation and partial nucleotide sequence of a cDNA clone for human histocompatibility antigen HLA-B by use of an oligodeoxynucleotide primer.
We have isolated a cDNA clone for one of the HLA-B locus alloantigens by hybridization with a 30-nucleotide-long DNA probe. The probe was isolated from a reverse transcriptase (RNA-dependent DNA nucleotidyltransferase)-catalyzed cDNA synthesis reaction on poly(A)-mRNA in which an oligonucleotide (5'-32P)dC-T-T-C-T-C-C-A-C-A-TOH served as a primer and in which dideoxynucleoside triphosphates were used to reduce the size and heterogeneity of the cDNA products. The desired cDNA clone was isolated from a library of recombinant cDNA clones in the plasmid pBR322. The partial nucleotide sequence of the cDNA clone corresponds to the amino acid sequence of HLA-B7 antigen. The approach described in this paper is extremely sensitive and may be useful in cloning other genes for which the corresponding mRNA is present at low levels. This cDNA clone is nearly full length and can be used to isolate and to study the genes within the HLA region and to obtain expression of HLA-B peptides in cells.
D001483:Base Sequence; D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D005796:Genes; D005813:Genes, Synthetic; D006680:HLA Antigens / Q000235:genetics*; D015235:HLA-B Antigens; D015793:HLA-B7 Antigen; D006801:Humans; D009841:Oligonucleotides / Q000235:genetics; D010957:Plasmids; D012333:RNA, Messenger / Q000235:genetics; D012194:RNA-Directed DNA Polymerase; D013698:Templates, Genetic
D004274:DNA, Recombinant; D006680:HLA Antigens; D015235:HLA-B Antigens; D015793:HLA-B7 Antigen; D009841:Oligonucleotides; D012333:RNA, Messenger; D012194:RNA-Directed DNA Polymerase