| 摘要 |
An object is to provide a TCR cloning system that enables not only bias-free analysis of TCR repertoires, but also collection of antigen-specific TCR ±/² cDNA pairs and evaluation of functions thereof. There is provided a method for producing a gene of T cell receptor (TCR) specific to an antigen A, which comprises 1) the step of stimulating a group of T cells including a T cell specific to an antigen A or one T cell specific to an antigen A under a condition effective for amplification of a TCR gene; 2) the step of identifying a T cell specific to an antigen A among the group of T cells including a T cell specific to the antigen A, and sorting one T cell specific to the antigen A into a vessel; and 3) the step of subjecting the one activated T cell specific to the antigen A in the vessel to PCR to amplify a gene of TCR specific to the antigen A. According to the present invention, a target TCR gene can be cloned within a shorter time compared with that required by the conventional methods, for example, about ten days. Further, according to the present invention, genes of TCR ± chain and ² chain can be highly efficiently cloned. Under the conditions of the examples, a pair of a TCR ± chain and TCR ² chain could be obtained from stimulated T cells sorted as single cells at a ratio of 100%. |
