Polymerase chain reaction with single-sided specificity: analysis of T cell receptor delta chain.
In the polymerase chain reaction (PCR), two specific oligonucleotide primers are used to amplify the sequences between them. However, this technique is not suitable for amplifying genes that encode molecules where the 5' portion of the sequences of interest is not known, such as the T cell receptor (TCR) or immunoglobulins. Because of this limitation, a novel technique, anchored polymerase chain reaction (A-PCR), was devised that requires sequence specificity only on the 3' end of the target fragment. It was used to analyze TCR delta chain mRNA's from human peripheral blood gamma delta T cells. Most of these cells had a V delta gene segment not previously described (V delta 3), and the delta chain junctional sequences formed a discrete subpopulation compared with those previously reported.
Loh
E Y
EY
Departments of Medicine and Microbiology and Immunology, Stanford University School of Medicine, CA 94305-5402.
Elliott
J F
JF
Cwirla
S
S
Lanier
L L
LL
Davis
M M
MM
D000595:Amino Acid Sequence; D001483:Base Sequence; D002460:Cell Line; D005784:Gene Amplification; D005796:Genes; D006801:Humans; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D012333:RNA, Messenger / Q000235:genetics; D012194:RNA-Directed DNA Polymerase; D011948:Receptors, Antigen, T-Cell / Q000235:genetics*; D013601:T-Lymphocytes / Q000276:immunology
D046911:Macromolecular Substances; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D011948:Receptors, Antigen, T-Cell; D012194:RNA-Directed DNA Polymerase