We report a set of testis-specific novel genes by comparing the The Institute for Genomic Research (TIGR) chicken gene index with the 13,132 expressed sequence tag (EST) collection from Korean Native Chicken (KNC). A total of 553 novel transcripts have resulted after comparison of our EST contigs and singlets against the chicken gene index and chicken genome sequences. Interestingly, 76% of the new genes were from testis-specific EST contigs or singlet and analysis of exon/intron structure showed that a significant number of the contigs have intronless coding regions. A large portion of the testis-specific transcripts have intronless coding region indicating that the testis-specific expression pattern is related to a group of single exon genes in the chicken. The chicken EST data that are described in this paper have been submitted to the NCBI dbEST under accession numbers CO759017-CO764274, CO765331-CO768903 and CO769672-CO773972.