中文摘要：

为了研究齐口裂腹鱼Schizothorax prenanti的低温适应性,根据齐口裂腹鱼卵巢转录组测序结果设计引物,采用RT-PCR法克隆得到齐口裂腹鱼zona pellucida 3（Schizothorax prenanti zona pellucida 3,sp-zp3）基因的编码区,将目的基因重组到表达载体,并将质粒转染至中国仓鼠的卵巢细胞（CHO-K1）中进行表达,分离纯化得到ZP3蛋白后,对其冰晶结合活性进行初步测定。结果表明：齐口裂腹鱼zp3基因在中国仓鼠卵巢细胞中高效表达,并纯化得到重组蛋白SP-ZP3,该蛋白具有一定的冰晶结合活性。研究表明,一些鱼类ZP蛋白可能具有冰晶结合活性,当选择压力存在时这些ZP蛋白将进化出具有冷适应性的功能。

英文摘要：

To understand the adaptation mechanism of Schizothorax prenanti to low temperature,we designed primers based on the RNA sequences of S.prenanti and amplified the full-length coding sequence of the Schizothorax prenanti zona pellucida 3（sp-zp3） by RT-PCR.The PCR products were cloned into the expression vector p IRES2-EGFP,which was transfected into Chinese hamster ovary（CHO-K1） cells,and then SP-ZP3 protein was obtained and purified.The highly purified SP-ZP3 protein was used in the ice-binding activity assay.The results indicated that zona pellucida 3 gene was expressed effectively in CHO-K1,and the recombinant protein was purified.Interestingly,the Zona pellucida 3 protein expressed in vitro exhibited ice-binding activity.These findings provide evidence indicating that some of fish ZP proteins may have ice-binding activity which can be evolved into cold-adaptive functions when selection pressures are present.