Molecular cloning of novel genes involved in T cell development using suppression subtractive hybridization

Objective: To clone and characterize novel genes which might be involved in T cell development. Methods: A subtracted cDNA library of mouse thymic stromal cell lines was constructed using suppression subtractive hybridization. RACE and RT PCR were used to clone and verify the full length cDNAs. Results: Differentially expressed gene fragments were obtained and sequencing revealed that 8 clones represented novel gene cDNAs, including C55 and C91.Northern blot analyses of C55 and C91 showed that the mRNA transcripts were 1.4 kb and 1.5 kb, respectively. Further, the expression level of C55 mRNA was significantly different between the two thymic stromal cell lines. The full length cDNAs of C55 and C91 have been successfully cloned and accepted by GenBank. Conclusion: suppression subtractive hybridization is an effective method to clone novel genes; Eight novel gene clones were obtained and two full length cDNAs cloned.