齐口裂腹鱼zona pellucida3的克隆表达及其蛋白活性研究  

Expression and protein activity of zona pellucida 3 from Schizothorax prenanti

作　　者：马战领 胡瑞芹 黄巧[1] 吴智超[1] 陈良标[1] 

机构地区：[1]上海海洋大学水产与生命学院水产种质资源发掘与利用省部共建教育部重点实验室,上海201306

出　　处：《大连海洋大学学报》2015年第6期592-597,共6页Journal of Dalian Ocean University

基　　金：国家基金委重点项目(31130049);上海市教委水产学一流学科项目(沪教科2012-62)

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。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.

关 键 词：齐口裂腹鱼 zona pellucida 3基因 冰晶结合活性 

分 类 号：S917.4[农业科学—水产科学]