| 菊黄东方鲀卵黄蛋白原肽段vWD的真核重组表达及功能研究 |
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| 引用本文： | 蒋彩云,乔琨,许旻,陈贝,刘智禹,黄文树.菊黄东方鲀卵黄蛋白原肽段vWD的真核重组表达及功能研究[J].集美大学学报(自然科学版),2021,26(4):299-305. |
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| 作者姓名： | 蒋彩云 乔琨 许旻 陈贝 刘智禹 黄文树 |
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| 作者单位： | ( 1.集美大学水产学院，福建 厦门 361021；2.福建省水产研究所，福建省海洋生物增养殖与高值化利用重点实验室，福建 厦门 361013；3.福建省海洋生物资源开发利用协同创新中心，福建 厦门 361021) |
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| 摘 要： | 河鲀的卵黄蛋白原可能参与河豚毒素（TTX）的体内转运过程。为了探讨该机理，建立了毕赤酵母真核表达系统，成功地表达了菊黄东方鲀（Takifugu flavidus）卵黄蛋白原的vWD结构域肽段（rTF_vWD），并采用Biacore-表面等离子体共振（SPR）系统检测了rTF_vWD与TTX的亲和力，以及腹腔注射小鼠验证rTF_vWD对TTX毒力的中和作用。结果表明:rTF_vWD与TTX的平衡解离常数（KD）为3.1 mmol/L；将TTX与rTF_vWD共孵后肌肉注射小鼠，2 h内小鼠的死亡率显著下降，表明TTX与rTF_vWD结合后，TTX的毒性降低了。
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| 关 键 词： | 河豚毒素 菊黄东方鲀 卵黄蛋白原肽段vWD 毕赤酵母 表面等离子体共振 中和作用 |
Eukaryotic Recombination Expression and Functional Study on Vitellogenin Peptide vWD from Takifugu flavidus |
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| JIANG Caiyun,QIAO Kun,XU Min,CHEN Bei,LIU Zhiyu,HUANG Wenshu.Eukaryotic Recombination Expression and Functional Study on Vitellogenin Peptide vWD from Takifugu flavidus[J].the Editorial Board of Jimei University(Natural Science),2021,26(4):299-305. |
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| Authors: | JIANG Caiyun QIAO Kun XU Min CHEN Bei LIU Zhiyu HUANG Wenshu |
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| Affiliation: | (1.Fisheries College,Jimei University,Xiamen 361021,China；2.Key Laboratory of Cultivation and High Value Utilization of Marine Organisms in Fujian Province,Fisheries Research Institute of Fujian,Xiamen 361013,China；3.Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources,Xiamen 361021,China) |
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| Abstract: | Vitellogenin of pufferfish may be involved in the in vivo transport of tetrodotoxin (TTX).In order to explore this mechanism,the eukaryotic expression system of the vitellogenin vWD domain peptides(rTF_vWD) of Takifugu flavidus in pichia pastoris was established and the target recombinant protein was successfully expressed in this study.The affinity between rTF_vWD and TTX was determined by using Biacore-surface plasma resonance(SPR) system.Intraperitoneal injection of mice was used to verify the neutralization of rTF_vWD and TTX in vivo.The results showed that the equilibrium dissociation constant (KD) of rTF_vWD and TTX was 3.1 mmol/L；When TTX and rTF_vWD were co-incubated and then intraperitoneally injected into mice,the mortality of mice within 2 h was significantly reduced,indicating that the toxicity of TTX was reduced combined with protein in vitro. |
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| Keywords: | tetrodotoxin Takifugu flavidus vitellogenin peptide vWD Pichia pastoris SPR Neutralization |
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