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.
作者
向勤
罗洁
吴金芋
肖朝新
李颜
樊汶樵
杨帆
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)
出处
《中国兽医杂志》
CAS
北大核心
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)
关键词
棘腹蛙
干扰素
克隆
序列分析
表达
Paa boulengeri
Interferon
Cloning
Sequence analysis
Expression
