Abstract
Sox (SRY-related HMG-box) genes encode a family of transcriptional regulators, which are characterized by a conserved 79-amino acid domain known as HMG-box. They play essential roles in a diverse range of processes including sex determination and the development of the central nervous system (CNS), neural crest and endoderm. In this paper, the HMG domain of ten distinct Sox gene family members (os-Sox2, os-Sox3a, os-Sox3b, os-Sox4, os-Sox11a, os-Sox11b, os-Sox14a, os-Sox14b, os-Sox21a, os-Sox21b) were isolated from both male and female Odorrana schmackeri (Boettger, 1892) using PCR, and no sexual differences were found. Molecular phylogenetic analysis of the HMG domain suggested that these ten Sox genes are members of the SoxB and SoxC groups. In addition, sequence analysis suggested that four Sox genes (os-Sox3, os-Sox11, os-Sox14, os-Sox21) were duplicated. The duplication-degeneration-complementation model should be implied to explain the evolution and diversity of the Sox gene family in O. schmackeri.
Highlights
Gene members belonging to the Sox family are characterized by a recognisable 240-nucleotide sequence that encodes a 79-amino acid motif known as the HMG-box domain
Many HMG-box genes act as transcription factors regulating gene expression during developmental patterning or cell differentiation
Sox1, Sox2, Sox3 are required for stem-cell maintenance in the central nervous system (CNS), and their effects are counteracted by Sox21 (SANDBERG et al 2005)
Summary
To isolate the HMG domain of the Sox genes, two male and female O. schmackeri were captured from Huangshan, Anhui. Total genomic DNA was obtained from muscle tissues with the Genomic DNA Extraction Kit (Axygen). A pair of degenerate primers were designed according to the sequence of the HMG-box in multiple Sox/SRY genes The PCR was carried out in a 25 μl reaction mixture containing 16μl ddH2O, 100 ng of genomic DNA, 1.5 mM Mg2+, 200 μM of each dNTP, 0.2 μM of each primer and 1 unit of Taq. DNA polymerase. The cycling conditions were 4 min at 95°C, followed by 5 cycles of 40s at 94°C, 40s at 48°C, 1 min 20 sec at 72°C 30 cycles of 40s at 94°C, 40s at 52°C, 1 min 20 sec at 72°C. The final extension was done during 10 min at 72°C
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