猪前胸腺肽α在大肠杆菌中的表达及其生物学活性鉴定  

Expression of porcine alpha-prothymosin in E.coli and its biological activity identification

作　　者：王錾彧 唐丽杰[1] 乔薪媛[1] 姜艳平[1] 崔文[1] 李一经[1] 

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨150030

出　　处：《中国预防兽医学报》2015年第1期53-56,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家科技支撑计划(2009BADB4B01)

摘　　要：为原核表达具有生物活性的重组前胸腺肽α(proTα),本实验利用人工合成的330 bp proTα DNA编码序列,构建重组表达质粒pET-proTα,将其转化于大肠杆菌Rosetta进行诱导表达,采用MTT方法测定重组蛋白对小鼠脾细胞和胸腺细胞增殖的影响。SDS-PAGE结果显示proTα重组蛋白以可溶形式表达,分子量约32 ku。重组蛋白经镍柱亲和层析纯化后获得具有生物活性的蛋白,在体外能够明显促进小鼠胸腺和脾淋巴细胞增殖活性,与轮状病毒共同免疫小鼠能够提高轮状病毒VP6蛋白的特异性抗体水平,并提高抗原刺激下脾细胞分泌IFN-γ水平。本研究首次确认了proTα蛋白在小鼠体内具有佐剂活性,为proTα生物活性的进一步研究及其作为疫苗佐剂的开发奠定基础。To express the porcine alpha-prothymosin （proTct） and identify its biological activity, the pET-proTα was constructed with a 330 bp synthesized porcine proTα gene and transformed into E.coli Rosetta for expression with IPTG induction. SDS-PAGE analysis indicated that the expressed recombinant protein of proTα was in a soluble form with a molecular weight about 32 ku. The proToα was purified by nickel affinity column, and the purified protein remarkably promoted the proliferation of mouse spleen and thymus lymphocytes, improved antibody responses of mouse to the VP6 of porcine rotavirus and enhanced the secreting of INF-γ in splenocyte. To our knowledge, this is the first report on the adjuvant activity of proTα for immune- enhancement in mice, and the data provided the basis for further research on proTα biological activity and its development as a vaccine adjuvant.

关 键 词：猪前胸腺肽α 大肠杆菌表达 生物学活性 

分 类 号：S852.61[农业科学—基础兽医学]