A consensus primer to amplify both alpha and beta chains of the human T cell receptor.
The use of reverse transcriptase in conjunction with the polymerase chain reaction (RT-PCR) has proven invaluable in the analysis of the T cell receptor (TCR) repertoire of different populations of T cells. However, the presence of a variable region in the T cell receptor has hindered the design of primers for the 5' end of the TCR cDNA. We describe the design and use of a degenerate consensus primer that allows amplification of both the alpha and beta chains of the human TCR. We have used this primer in the analysis of the TCR distribution of T cell clones, peripheral blood lymphocytes and lymphocytes residing in tissue. In addition, the primer has allowed the identification of an alternative splice site in the beta chain constant region which cannot translate into a functional constant region. We have found the primer to be easy to use, sensitive and specific.
Moonka
D
D
Department of Medicine, University of Pennsylvania Medical Center, Philadelphia.
Loh
E Y
EY
D017398:Alternative Splicing / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002999:Clone Cells; D016384:Consensus Sequence; D017931:DNA Primers; D018076:DNA, Complementary / Q000096:biosynthesis; D004587:Electrophoresis, Agar Gel; D005784:Gene Amplification; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012313:RNA / Q000302:isolation & purification; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics* / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology
D017931:DNA Primers; D018076:DNA, Complementary; D016693:Receptors, Antigen, T-Cell, alpha-beta; D012313:RNA