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日本血吸虫蛋白酶体α2亚基基因的克隆、表达及功能分析
引用本文:洪炀,韩宏晓,彭金彪,李晔,石耀军,傅志强,刘金明,李祥瑞,林矫矫.日本血吸虫蛋白酶体α2亚基基因的克隆、表达及功能分析[J].生物工程学报,2010,26(4):509-516.
作者姓名:洪炀  韩宏晓  彭金彪  李晔  石耀军  傅志强  刘金明  李祥瑞  林矫矫
作者单位:1. 中国农业科学院上海兽医研究所,农业部动物寄生虫学重点开放实验室,上海,200241;南京农业大学动物医学院,南京,210095
2. 中国农业科学院上海兽医研究所,农业部动物寄生虫学重点开放实验室,上海,200241
3. 南京农业大学动物医学院,南京,210095
基金项目:国家自然科学基金 (No. 30671581),国家重点基础研究发展计划项目 (973计划) (No. 2007CB513108),国家高技术研究发展计划 (863计划) (No. 2006AA10A207),国家科技支撑计划 (No. 2006BAD06A09) 资助。
摘    要:26S蛋白酶体是一种能够降解大多数内源性蛋白的多亚基复合物,它的蛋白降解作用能够影响细胞周期、转录控制和其他一些重要的细胞进程。本实验利用PCR技术从日本血吸虫18d童虫中首次扩增到蛋白酶体α2亚基基因(GenBank Accession No.AY813725),序列分析表明该基因的开放阅读框(ORF)含708bp,编码235个氨基酸,理论分子量25.84kDa。同源性分析结果显示,该基因为日本血吸虫蛋白酶体α2亚基,命名为SjPSMA2。实时定量PCR分析显示该基因在7d、13d、18d、23d、32d和42d虫体中都有表达,7d和23d虫体表达量低于其他几个时期。构建了该基因的原核表达质粒pET28a(+)-SjPSMA2,在大肠杆菌系统中成功获得了表达,重组蛋白以包涵体形式存在,Western blotting显示表达产物能被日本血吸虫成虫粗抗原免疫血清所识别,并且能检测到天然状态下该蛋白的存在。应用重组蛋白免疫BALB/c小鼠后,诱导产生了较高的特异性抗体水平及12.33%的减虫率和35.23%的肝脏减卵率。SjPSMA2基因及其表达产物的获得,为探索蛋白酶体在血吸虫生长发育中的作用提供了重要基础。

关 键 词:日本血吸虫  蛋白酶体α2亚基  克隆  表达  免疫保护
收稿时间:2009/12/16 0:00:00

Cloning,expression and characterization of a gene encoding α2 subunit of the proteasome in Schistosoma japonicum
Yang Hong,Hongxiao Han,Jinbiao Peng,Ye Li,Yaojun Shi,Zhiqiang Fu,Jinming Liu,Xiangrui Li and Jiaojiao Lin.Cloning,expression and characterization of a gene encoding α2 subunit of the proteasome in Schistosoma japonicum[J].Chinese Journal of Biotechnology,2010,26(4):509-516.
Authors:Yang Hong  Hongxiao Han  Jinbiao Peng  Ye Li  Yaojun Shi  Zhiqiang Fu  Jinming Liu  Xiangrui Li and Jiaojiao Lin
Institution:Key Laboratory of Animal Parasitology, Ministry of Agriculture, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Science, Shanghai 200241, China; College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China;Key Laboratory of Animal Parasitology, Ministry of Agriculture, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Science, Shanghai 200241, China; College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China;Key Laboratory of Animal Parasitology, Ministry of Agriculture, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Science, Shanghai 200241, China;Key Laboratory of Animal Parasitology, Ministry of Agriculture, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Science, Shanghai 200241, China;Key Laboratory of Animal Parasitology, Ministry of Agriculture, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Science, Shanghai 200241, China;Key Laboratory of Animal Parasitology, Ministry of Agriculture, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Science, Shanghai 200241, China;Key Laboratory of Animal Parasitology, Ministry of Agriculture, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Science, Shanghai 200241, China;College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China;Key Laboratory of Animal Parasitology, Ministry of Agriculture, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Science, Shanghai 200241, China
Abstract:
Keywords:Schistosoma japonicum  proteasome α2 subunit (PSMA2)  clone  expression  immunoprotection
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