Paired cloning of the T cell receptor alpha and beta genes from a single Tcell without the establishment of a T cell clone

T cell receptors, which recognize antigen peptides on MHC molecules, are es sential probes for the analysis of T cell antigen specificity. The identifi cation of paired T cell receptor (TCR) chains, alpha/beta or gamma/delta, u sually requires the establishment of T cell clones, which is not always ava ilable. In this study, we tried, as an alternative method, the paired cloni ng of TCR alpha/beta genes directly from a single T cell. T cells were sort ed as a single cell from which RNA was extracted. Then, TCR alpha/beta CDR3 regions were amplified from the single cell-derived cDNA by reverse transc riptase-polymerase chain reaction to determine their sequences. We successf ully identified pairs of TCR alpha/beta genes, and reconstructed the TCR mo lecule by a bacterial expression system. This strategy makes it possible to obtain recombinant TCR molecules from a single T cell without cellular clo ning and promotes the investigation of T cell antigen specificity.