| 鸡白细胞介素18成熟蛋白在昆虫杆状病毒系统中的表达及其活性检测 |
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| 引用本文： | 范忠玲,王婷婷,马凤龙,胡敬东. 鸡白细胞介素18成熟蛋白在昆虫杆状病毒系统中的表达及其活性检测[J]. 山东农业大学学报(自然科学版), 2011, 42(3): 438-444 |
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| 作者姓名： | 范忠玲 王婷婷 马凤龙 胡敬东 |
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| 作者单位： | 1. 山东农业大学动物科技学院,山东泰安,271018
2. 济南军区疾病预防控制中心,山东济南,250014 |
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| 基金项目： | 山东省优秀中青年科学家科研奖励基金(2006BS06015); 山东省自然科学基金(Y2008D12)资助 |
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| 摘 要： | 首先将鸡白细胞介素18成熟蛋白(mature chicken interleukin-18,mChIL-18)基因亚克隆至杆状病毒转移载体pFastBac HTb上,然后转化至含穿梭载体Bacmid的受体菌E.coli DH10BacTM中,构建重组Bacmid(rBacmid)。通过脂质体介导法将纯化的rBacmid转染sf9细胞,获得完整重组杆状病毒,将达到一定滴度的重组杆状病毒感染sf9,收获感染后不同时间段的培养上清和细胞,经SDS-PAGE分析、Western-blotting和间接免疫荧光(IFA)检测,结果表明,分子量约为23KDa的重组蛋白在昆虫细胞中获得了表达;鸡淋巴细胞转化试验和水疱性口炎病毒(VSV)抑制试验表明,表达产物具有良好的生物学活性。结论:在杆状病毒表达系统中成功表达了有活性的mChIL-18蛋白。
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| 关 键 词： | ChIL-18成熟蛋白 杆状病毒表达载体系统 昆虫细胞 生物学活性 |
EXPRESSION OF CHICKEN INTERLEUKIN-18 MATURATION PROTEIN GENE IN INSECT CELLS AND IDENTIFICATION OF BIOACTIVITY OF ITS EXPRESSED PROTEIN |
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| FAN Zhong-ling,WANG Ting-ting,MA Feng-long,HU Jing-dong. EXPRESSION OF CHICKEN INTERLEUKIN-18 MATURATION PROTEIN GENE IN INSECT CELLS AND IDENTIFICATION OF BIOACTIVITY OF ITS EXPRESSED PROTEIN[J]. Journal of Shandong Agricultural University, 2011, 42(3): 438-444 |
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| Authors: | FAN Zhong-ling WANG Ting-ting MA Feng-long HU Jing-dong |
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| Affiliation: | FAN Zhong-ling1,WANG Ting-ting1,MA Feng-long2,HU Jing-dong1*(1.College of Animal Science & Veterinary Medicine,Shandong Agricultural University,Tai'an,271018,China,2.The Center for Disease Control and Prevention of Jinan Military Area of PLA,Shandong,Jinan,250014,China) |
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| Abstract: | To obtain the recombinant Bacmid,the mature chicken interleukin-18 protein(mChIL-18) gene was subcloned into the baculovirus transfer vector pFastBac HTb and the recombinant plasmid was transformed into competent DH10BacTM E.colicells containing bacu-lovirus shuttle vector bacmid.Then the purified recombinant bacmid was transfected into sf9 insect cells by the method of using lipofectin for producing integrated recombinant baculo-virus.Infected the sf9 with the higher baculoviral stock for expressing protei... |
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| Keywords: | ChIL-18 maturate protein baculovirus express system insect cells bioactivity |
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