NOWOSLAWSKI, L,; MURDOCK, C.A.; WIBBELS, T.*; University of Alabama at Birmingham; University of Alabama at Birmingham; University of Alabama at Birmingham: PCR-based strategy for cloning and development of a quantitative competitive RT-PCR for SOX9 in a turtle with temperature-dependent sex determination.

A variety of genes have been identified which may be involved in sex determination and sexual differentiation of vertebrates. Some of these genes appear to be conserved among vertebrates. SOX9 has been identified in a variety of vertebrates and it has been suggested to be involved in testicular differentiation. In the current study, a PCR-based strategy was developed in an attempt to clone SOX9 from a turtle, Trachemys scripta, with temperature-dependent sex determination. Initially a 961 bp fragment was successfully amplified. The data from that fragment was used to generate a competitor fragment which contained an approximate 83 bp deletion. We are currently using 5′ and 3′ RACE to clone the entire cDNA for phylogenetic comparisons to those of other vertebrates. Additionally, we will utilize the competitor in a quantitative competitive RT-PCR to precisely quantify the expression of SOX9 during sex determination and sexual differentiation in this turtle.