Background: The origin of functional innovation is among the key questions in biology. Recently, it has been shown that new genes could arise from non-coding DNA and that such novel genes are often involved in male reproduction.
Results: With the aim of identifying novel genes, we used the technique "generation of longer cDNA fragments from serial analysis of gene expression (SAGE) tags for gene identification (GLGI)" to extend 84 sex-biased 3'end SAGE tags that previously could not be mapped to the D. pseudoobscura transcriptome. Eleven male-biased and 33 female-biased GLGI fragments were obtained, of which 5 male-biased and 3 female-biased tags corresponded to putatively novel genes. This excess of novel genes with a male-biased gene expression pattern is consistent with previous results, which found novel genes to be primarily expressed in male reproductive tissues. 5' RACE analysis indicated that these novel transcripts are very short in length and could contain introns. Interspecies comparisons revealed that most novel transcripts show evidence for purifying selection.
Conclusion: Overall, our data indicate that among sex-biased genes a considerable number of novel genes (approximately 2-4%) exist in D. pseudoobscura, which could not be predicted based on D. melanogaster gene models.