棘腹蛙Ⅰ型干扰素基因的克隆与原核表达  

Clone and expression of type Ⅰ interferon from Paa boulengeri

作　　者：向勤[1] 罗洁[1] 吴金芋 肖朝新 李颜[1] 樊汶樵[1] 杨帆[1] XIANG Qin LUO Jie WU Jin-yu XIAO Chao-xin LI Yan FAN Wen-qiao YANG Fan(College of Forestry ＆ Life Science Chongqing University of Art ＆ Science Chongqihg Chongqing Research Centers of Conservation and Development on Rare ＆ Endangered Aquatic Resources, Chongqing 402168, China)

机构地区：[1]重庆文理学院林学与生命科学学院重庆珍稀濒危水产资源保护与开发研究中心,重庆永川402168

出　　处：《中国兽医杂志》2017年第4期23-26,共4页Chinese Journal of Veterinary Medicine

基　　金：国家级大学生创新创业训练计划项目(201510642064);重庆市科技计划重点专项(cstc2015jcyjBX0013);重庆市科技计划项目(cstc2014jcyjA80042);重庆文理学院人才引进项目(R2013LS13;R2014LX07)

摘　　要：干扰素(IFN)具有抗病毒、抗肿瘤和免疫调节功能,具备较好的临床应用前景。为了解棘腹蛙干扰素的基因信息,本试验明确了棘腹蛙Ⅰ型IFN基因ORF编码长度为561 bp,编码的蛋白质含有1个信号肽和103aa的保守功能结构域。遗传进化分析显示,棘腹蛙Ⅰ型IFN进化相对保守;其蛋白质结构主要由α螺旋构成,细胞表面受体结合能力相对保守。将目的片段装入原核表达载体pET32a并转化BL21(DE3)进行蛋白表达与纯化,获得了预期蛋白。本研究证实了棘腹蛙Ⅰ型IFN属于相对保守的典型Ⅰ型干扰素;为后期深入挖掘棘腹蛙IFN的生物学功能、促进广谱抗病毒生物制剂研发提供了参考。Interferons have the functions of anti-virus, anti-tumor and immune regulation. For better understanding of the interferon encoding genes, we have studied the full length sequence of type I interferon from Paa boulengeri by RT-PCR. The results showed that, IFN-Pb, was 649bp long, encoding 186 aa protein and containing a signal peptide and a conserved interferon functional domain, especially consisting of an alpha helix composition and a cell surface receptor binding domain. Besides, phylogenic analysis illustrated that it had a closer relationship with Xenopus, and was clustered into aquatic branches. The IFN-Pb was inserted into pro- karyotic expression vector pET32a （ ＋ ） by restriction enzyme digestion and then the plasmids was transformed in E. coli BL21 （DE3）. The protein was induced with IPTG and the fusion protein was purified successfully with Ni-sepahrose.

关 键 词：棘腹蛙 干扰素 克隆 序列分析 表达 

分 类 号：S947[农业科学—水产养殖]