WIBBELS, T.; MURDOCK, C.; ZELICKSON, M.; SPARKS, K.: Development of a quantitative competitive RT-PCR for Dmrt1 in a turtle with temperature-dependent sex determination.

The Dmrt1 gene appears to be involved in gonadal development in a variety of vertebrates. Expression patterns in mammals suggest that this gene could be involved in male sex determination and/or testicular differentiation. In the current study, a quantitative competitive RT-PCR (QC-RT-PCR) was developed to examine the expression of Dmrt1 during temperature-dependent sex determination in the red-eared slider turtle, Trachemy scripta. Initially, a fragment of turtle Dmrt1 cDNA was isolated by PCR and then 5′ and 3′ RACE were used to clone the full length cDNA. Based on the cDNA structure, primers were designed to generate a 384 base pair fragment of the Dmrt1 cDNA which included an 84 base pair deletion. That fragment was then used to develop a QC-RT-PCR for turtle Dmrt1. The availability of a QC-RT-PCR provides a highly sensitive means of examining the expression of Dmrt1 during temperature-dependent sex determination.