棘腹蛙Ⅰ型干扰素基因的克隆与原核表达

发布时间：2018-11-11 19:21

【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。
[Abstract]:Interferon (IFN) has antiviral, antitumor and immunomodulatory functions, and has a good clinical application prospect. In order to understand the gene information of IFNs, the protein encoded by 561 bp, ORF of the type I IFN gene of Rana spinosus contained a signal peptide and a conserved functional domain of 103aa. Genetic evolution analysis showed that the evolution of type I IFN was relatively conserved, and its protein structure was mainly composed of 伪 helix, and the receptor binding ability on cell surface was relatively conserved. The target fragment was inserted into prokaryotic expression vector pET32a and transformed into BL21 (DE3) for protein expression and purification. This study confirmed that the type I IFN belongs to the typical type I interferon which is relatively conserved, which provides a reference for further exploring the biological function of IFN and promoting the development of broad-spectrum antiviral biological agents.
【作者单位】：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;
【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)
【分类号】：Q78;S917.4

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