文章摘要
黄颡鱼细胞因子信号传导抑制因子SOCS1的原核表达及多克隆抗体制备
Prokaryotic protein expression of suppressor of cytokine signaling 1 of yellow catfish Pelteobagrus fulvidraco, and preparation of their polyclonal antibodies
投稿时间:2019-06-23  修订日期:2019-09-18
DOI:
中文关键词: 黄颡鱼  SOCS1  原核表达  多克隆抗体制备  蛋白表达
英文关键词: Pelteobagrus fulvidraco  SOCS1  prokaryotic expression  preparation of polyclonal antibody  protein expression
基金项目:(31572605)和“蓝色粮仓”科技创新专项(2018YFD0900400)。
作者单位E-mail
叶汉梅 华中农业大学水产学院 1447394651@qq.com 
魏晓雷 华中农业大学水产学院  
谭肖英 华中农业大学水产学院 txy7933@163.com 
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中文摘要:
      细胞因子信号传导抑制因子(suppressor of cytokine signaling, SOCS)是一类由细胞产生的胞内蛋白,能够反馈性阻断细胞因子信号传导过程的负性调控因子。为深入研究黄颡鱼细胞因子信号传导的发生过程和机制,本研究从黄颡鱼cDNA中克隆获得561bp的黄颡鱼SOCS1(PfSOCS1)基因编码序列,成功构建了重组质粒pET32a(+)-PfSOCS1,并转化至大肠杆菌 BL21(DE3)感受态细胞中,在不同温度、IPTG 浓度、诱导时间等条件下进行表达,确定最佳表达条件。本实验采用包涵体纯化的方法得到纯度较高的重组SOCS1蛋白;以重组PfSOCS1蛋白免疫Balb/C小鼠,制备多克隆抗体。通过Western blot鉴定了PfSOCS1抗血清可以特异性识别重组PfSOCS1蛋白。蛋白水平的组织分布显示,黄颡鱼SOCS1蛋白在肝脏中表达水平较高。
英文摘要:
      Suppressor of cytokine signaling (SOCS) is a class of intracellular proteins that inhibited cytokine signaling in. We cloned the sequence of SOCS1 gene from yellow catfish Pelteobagrus fulvidraco, whose length was 561bp. We constructed the recombinant expression plasmid [pET32a (+)-SOCS1]Scarrying PfSOCS1 protein. The recombinant PfSOCS1 protein was optimally expressed by transformation into Escherichia coli BL21 (DE3) and then highly purified under inclusion body conditions. The polyclonal anti-PfSOCS1 antibodies were prepared by immunizing Balb/C mouse with recombinant PfSOCS1 protein, which could be used to recognize recombinant PfSOCS1 protein by Western blotting. The protein expression of PfSOCS1 was detected in kidney, intestine, liver and ovary, with relatively predominant level in the liver in P. fulvidraco.
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