乌龟Sox gene的PCR扩增及其条件

摘要

The sox gene of Chinemys reevessi was amplified by using a pair of primers which could amplify the conservative motif of human SRY gene. The amplification band with the length of 220bp was observed in both male and female Chinemys,which was consistent with that of human SRY gene. The optimization system of PCR reaction was 2.0 mM Mg2+,dNTP 120μM,0.3μM primer and annealing at 52℃. This research presents molecular proof for the mechanism of turtle's sex determination and the conservation of sox gene's evolution.%参照人SRY gene HMG-box保守区的序列，设计一对引物，扩增了乌龟的Sox gene,并对扩增条件进行了优化。结果显示乌龟Sox gene的扩增片段与人SRY gene 扩增片断大小相同，为220bp左右，且雌雄个体间无差异；最佳扩增条件为：Mg2＋2.0mM、dNTP 120μM、引物0.3μM及退火温度52℃。本研究为探索乌龟的性别决定机制以及Sox gene进化的保守性提供分子资料。