| Bac-to-Bac/BmNPV杆状病毒在家蚕幼虫中表达重组鲎C因子 |
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| 引用本文： | 祁静,刘涛,李珍,贡成良,吴海苹,张春.Bac-to-Bac/BmNPV杆状病毒在家蚕幼虫中表达重组鲎C因子[J].生物工程学报,2014,30(10):1594-1601. |
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| 作者姓名： | 祁静 刘涛 李珍 贡成良 吴海苹 张春 |
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| 作者单位： | 1 中国科学院苏州生物医学工程技术研究所 苏州市分子诊断与治疗技术重点实验室，江苏 苏州 215163;1 中国科学院苏州生物医学工程技术研究所 苏州市分子诊断与治疗技术重点实验室，江苏 苏州 215163;2 苏州大学基础医学与生命科学学院，江苏 苏州 215006;2 苏州大学基础医学与生命科学学院，江苏 苏州 215006;3 厦门市鲎试剂实验厂有限公司，福建 厦门 361022;1 中国科学院苏州生物医学工程技术研究所 苏州市分子诊断与治疗技术重点实验室，江苏 苏州 215163 |
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| 基金项目： | 苏州市科技局项目 (No. ZXY2012011) 资助。 |
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| 摘 要： | 鲎C因子是鲎血细胞中一种对内毒素具有高亲和力的丝氨酸蛋白酶,被内毒素激活后可水解人工合成的三肽底物,因而能替代传统的鲎试剂用于内毒素检测。通过RT-PCR从东方鲎血细胞中扩增出鲎C因子基因(factor C)序列,利用Bac-to-Bac/BmNPV杆状病毒表达系统在家蚕幼虫中表达该蛋白,并对其进行活性测定。结果显示:被感染的家蚕血清稀释后能检测到Factor C活性,稀释500倍的血清可以用于内毒素检测,且其灵敏度能达到0.2 EU/mL,有望开发新型的、低成本的内毒素检测试剂。
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| 关 键 词： | 鲎C因子 家蚕幼虫 Bac-to-Bac/BmNPV杆状病毒表达系统 内毒素检测试剂 |
| 收稿时间： | 2013/12/25 0:00:00 |
Expression of limulus Factor C in silkworm larvae by Bac-to-Bac/BmNPV baculovirus expression system |
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| Jing Qi,Tao Liu,Zhen Li,Chengliang Gong,Haiping Wu and Chun Zhang.Expression of limulus Factor C in silkworm larvae by Bac-to-Bac/BmNPV baculovirus expression system[J].Chinese Journal of Biotechnology,2014,30(10):1594-1601. |
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| Authors: | Jing Qi Tao Liu Zhen Li Chengliang Gong Haiping Wu and Chun Zhang |
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| Institution: | 1 Suzhou Municipal Key Laboratory of Molecular Diagnostics and Therapeutics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou 215163, Jiangsu, China;1 Suzhou Municipal Key Laboratory of Molecular Diagnostics and Therapeutics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou 215163, Jiangsu, China;2 School of Biology and Basic Medical Science, Soochow University, Suzhou 215006, Jiangsu, China;2 School of Biology and Basic Medical Science, Soochow University, Suzhou 215006, Jiangsu, China;3 Chinese Horseshoe Crab Reagent Manufactory, Co., Ltd., Xiamen 361022, Fujian, China;1 Suzhou Municipal Key Laboratory of Molecular Diagnostics and Therapeutics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou 215163, Jiangsu, China |
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| Abstract: | Limulus Factor C, a serine protease zymogen from the amoebocytes of the limulus, has high affinity for endotoxin. When Factor C is activated by endotoxin, it hydrolyses artificial tripeptide substrate and measurable products are released, so it can be used as an alternative reagent for endotoxin analysis. Factor C gene of Tachypleus tridentatus was obtained through RT-PCR and the recombinant protein was expressed by Bac-to-Bac/BmNPV baculovirus expression system in silkworm larvae. The activity of Factor C was detected with diluted serum of silkworm larvae , and the sensitivity of endotoxin detected was 0.2 EU/mL when the serum was diluted at 1:500. The silkworm larvae expressed limulus Factor C could be used to develop a new low-cost endotoxin test reagent. |
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| Keywords: | limulus Factor C silkworm larvae Bac-to-Bac/BmNPV baculovirus expression system endotoxin test reagent |
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