赤链蛇不同组织Sox基因表达的RT-PCR分析

The RT-PCR Analysis on the Sox Gene Expression of Different Tissues from the Dinodon rufozonatum

采用RT PCR技术,研究了赤链蛇不同组织Sox基因的表达。通过PCR产物直接克隆法和SSCP技术筛选阳性克隆,分析了雄性睾丸和雌性卵巢组织中的Sox基因序列。结果显示,在赤链蛇雌雄成体组织中,Sox基因在睾丸、卵巢、脑和脾组织中均有不同程度的表达,而在雌雄成体肌肉组织中均无表达,显示该基因表达有一定的组织特异性。序列分析显示睾丸组织中表达的是DRSox3,卵巢组织中表达的是DRSox22。在Sox家族中,Sox3表达于中枢神经系统和尿生殖嵴的发育过程中;Sox22则表达于多种组织和神经系统中,可横跨CNS和PNS的整个过程。此结果表明Sox基因不仅在性别决定中起作用,还可能在胚胎发育过程中担负重要功能。

The Sox gene expression in different tissues from the Dinodon rufozonatum was studied by using RT-PCR technique,and the Sox gene fragments from the testicle and ovary were cloned using RT-PCR products. The positive clone were identified by SSCP and sequenced. The results show that the Sox gene has specific expression in testicle,ovary,brain,spleen,and do not express in muscle for both male and female sample. The sequence analysis reveal that the expressed Sox genes in testicle is DRSox3 and in ovary is DRSox22,which suggested that there are difference between the sex gland of male and female tissues. As already mentioned in the Sox family,Sox3 is expressed in the developing central nervous system and also expressed in somatic cells of the early urogenital ridge; Sox22 is expressed throughout embryonic CNS and PNS,and is also found expressing in developing mesenchymal structures,fetal kidney,heart,pancreas and gonads. This support that the Sox gene act an important role not only on the sex determination, but also on the developmental processes of early embryogenesis. The results provided molecular data for the study of embryogenesis of Dinodon rufozonatum.