Studies On The Sox Gene Of Rana Rugulosa Wiegmann

This paper consists of two parts, which are as follows:PART ONE: reviews;The progress in research on sex determining genes was explained on the clue of SRY gene. Sex determination involved a cascade of events initiated by the SRY gene. Apart from SRY, other genes, such as SOX gene family ,MIS,SF-1,DAX1,DSS also take part in sex determination.PART TWO: studiesl.The Sox gene of Rana rugulosa Wiegmann was amplified by using the technique of PCR with a pair of primers which specially amplify the conserved motif (HMG-box) of human SRY gene. There was only a band, about 221bp,in both male and female Rana rugulosa Wiegmann .The optimization system of PCR reaction was 2.0mM Mg2+, 90nMdNTP, 0.4uM primer and annealing at 54.2.Using a pair of degenerate primers based on the conserved region, HMG-box, of human SRY gene, eight different fragments were amplified from both female and male Rana rugulosa Wiegmann then cloned by using pMD18-T vector and sequenced. The result indicated that they showed high homology to Human SOX genes, and the identities to human SOX genes in the DNA sequence and the amino acid sequence were 87% ?68%, 88%, 83%, 88% and 92%, 76%, 94%, 98%, 98%, respectively, which may be concluded that Sox genes were highly conservative in phylogenetic.3.The expression of Sox genes in different tissue of Rana rugulosa Wiegmann was analysed by using RT-PCR technique. An expression was observed in testis ,brain ,heart ,and ovaries, but no expression was found in tissue of liver and muscle. The results showed that the expression of Sox genes had some tissue speciality.Our work provides molecular data for the study of sex-determining mechanism of Rana rugulosa Wiegmann, which may be temperature-dependent sex determination, and offers the proof for the conservation of Sox genes' evolution.