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-《集美大学学报（自然科学版）》

文章信息/Info

Title:: Eukaryotic Recombination Expression and Functional Study on Vitellogenin Peptide vWD from Takifugu flavidus

作者:: 蒋彩云1; 乔琨2; 许旻2; 陈贝2; 刘智禹2; 3; 黄文树1; 3; ( 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)

Author(s):: JIANG Caiyun1; QIAO Kun2; XU Min2; CHEN Bei2; LIU Zhiyu2; 3; HUANG Wenshu1; 3; (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)

Keywords:: tetrodotoxin; Takifugu flavidus; vitellogenin peptide vWD; Pichia pastoris; SPR; Neutralization

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。

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.