梅花鹿IFN-β1的原核表达及活性鉴定  

Prokaryotic expression of interferon-bata1 of sika deer and the activity identification

作　　者：苏凤艳[1] 李哲[1] 刘存发 宗颖[1] 曾范利[1] 王全凯[1,3] 

机构地区：[1]吉林农业大学中药材学院,吉林长春130118 [2]吉林省野生动物救护繁育中心,吉林长春130122 [3]吉林省中韩动物科学研究院,吉林长春130600

出　　处：《中国预防兽医学报》2015年第2期136-139,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：吉林省自然科学基金(201115194);国家国际科技合作专项(2011DFA32900)

摘　　要：为表达具有抗病毒活性的梅花鹿干扰素β1(IFN-β1),本研究通过PCR扩增梅花鹿IFN-β1基因,将其克隆至原核表达载体p ET-28a中,构建重组质粒p ET-IFNβ1,并将其转化至大肠杆菌Rosetta(DE3)感受态细胞中,经IPTG诱导后,得到高效表达。SDS-PAGE与western blot分析结果表明,梅花鹿IFN-β1重组蛋白的分子量大小约为24 ku,表达产物主要为不溶性的包涵体,表达量占菌体总蛋白的59.02%。将Ni柱纯化的p ET-IFNβ1诱导表达产物复性后,细胞病变抑制法检测表明表达产物具有抗病毒活性,其抗病毒活性可以达到10×53.81U/0.1 m L。梅花鹿INF-β1的表达为梅花鹿INF-β1的应用奠定了基础。To express the sika deer interferon batal （IFN-β1）, the IFN-β1 gene was amplified by PCR from genome DNA extracted from liver of sika deer, cloned and sequenced. The results showed the sequence of sika deer IFN-β1 gene was 498 bp, which encodes 165 amino acid residues. Then, the IFN-β1 gene was inserted into pET-28a vector to construct recombinant plasmid of pET-IFNβ1, pET-IFNβ1 was transformed into E.coli Rosseta （DE3）. The recombinant IFN-β1 （rIFN-β1） was expressed by IPTG inducing. The expressed rIFN-β1 was about 24 ku and mainly in the form of inclusion body which was account for 59.02% of the total bacteria protein. The rIFN-β1 was further purified by the Ni column and renatured. Then, cytopathic inhibition method detected that rIFN-β1 possess antiviral activity which was about 10×5^3.82 U/0.1 mL. This study provided a basis for further study and application of sika deer IFN-β1.

关 键 词：梅花鹿 IFN-β1基因 原核表达 活性鉴定 

分 类 号：S858.94[农业科学—临床兽医学]