[1]刘文强,刘桂芹,路建彪,等.羊白细胞介素-18成熟蛋白的酵母表达和生物学活性研究[J].中国预防兽医学报,2009,(12):969-973.
 LIU Wen-qiang,LIU Gui-qin,LU Jian-biao,et al.Expression of goat IL-18 mature protein in Pichia mammalian expression system[J].Chinese journal of preventive veterinary medicine,2009,(12):969-973.
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羊白细胞介素-18成熟蛋白的酵母表达和生物学活性研究
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2009年12期
页码:
969-973
栏目:
免疫学
出版日期:
2009-12-15

文章信息/Info

Title:
Expression of goat IL-18 mature protein in Pichia mammalian expression system
文章编号:
1008-0589(2009)12-0969-05
作者:
刘文强1 刘桂芹1路建彪1赵宏坤2许传田3
1. 聊城大学 农学院,山东 聊城 252059;
2. 山东农业大学 动物科技学院,山东 泰安 271018;
3. 山东省农科院畜牧所,山东 济南250100
Author(s):
LIU Wen-qiang1 LIU Gui-qin1 LU Jian-biao1 ZHAO Hong-kun2 XU Chuan-tian3
1. College of Agriculture of Liaocheng University, Liaocheng 252059, China;
2. College of Animal Science and Technology of Shandong Agricultural University, Taian 271018, China;
3. Institute of Animal Science and Veterinary Medicine Shandong Agricultural Sciences, Jinan 250100, China
关键词:
山羊白细胞介素-18酵母表达系统活性
Keywords:
goat IL-18 yeast expression system bioactivity
分类号:
S852.4
文献标志码:
A
摘要:
为了研究重组羊白细胞介素(gIL)-18在酵母系统中的高效表达,进一步阐明该重组蛋白的生物学活性,以含有gIL-18基因的重组质粒为模板进行PCR扩增,构建重组表达质粒pPICZ-gIL-18,转化于毕赤酵母GS115,以甲醇诱导表达,经SDS-PAGE和western blot分析证实了重组蛋白的表达,分泌的重组gIL-18表达量为100 mg/L。经纯化后用MTT法和MDBK-VSV法检测表达的重组IL-18体外生物活性,利用免疫试验检测了重组蛋白对羊痘疫苗的免疫增强作用。实验结果表明,该蛋白具有诱导MDBK细胞分泌IFN-γ和刺激PBMC增殖的生物学活性,比活性为1.5×105 u/mg。体外具有增强羊痘疫苗的活性。毕赤酵母分泌表达的重组gIL-18具有良好的生物学活性,为gIL-18作为免疫佐剂和免疫治疗剂的大规模应用奠定了基础。
Abstract:
For expression of goat interleukin-18 (gIL-18) in pichia pastoris system, the IL-18 gene was amplified by PCR and cloned into pPICZαA vectors, then transformed into pichia pastoris system GS115. High level expression of recombinant protein up to 100 mg/L was achieved following inducing with methanol. Bioactivity of recombinant protein was analyzed by MTT and MDBK-VSV methods. The enhanced immunity was measured by goat pox vaccine test. The recombinant protein could stimulate PBMCs proliferation and induce IFN-γ production in MDBK cells. It also showed immune enhancement to the antigen of goat pox vaccine. The recombinant gIL-18 expressing in yeast system retained nature bioactivity and could be a valuable resource for the large-scale clinical use as an immune adjuvant or therapeutic cytokines.
  

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备注/Memo

备注/Memo:

收稿日期:2009-04-06
基金项目:国家自然科学基金(30800130);山东省中青年科学家奖励基金(2006BS02012)
作者简介:刘文强(1977-),男,山东临清人,博士,副教授,研究方向为动物传染病学.



更新日期/Last Update: 2010-06-11