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  

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

作　　者：蒋彩云 乔琨[2] 许旻[2] 陈贝[2] 刘智禹[2,3] 黄文树 JIANG Caiyun;QIAO Kun;XU Min;CHEN Bei;LIU Zhiyu;HUANG Wenshu(Fisheries College,Jimei University,Xiamen 361021,China;Key Laboratory of Cultivation and High-Value Utilization of Marine Organisms in Fujian Province,Fisheries Research Institute of Fujian,Xiamen 361013,China;Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources,Xiamen 361021,China)

机构地区：[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

出　　处：《集美大学学报（自然科学版）》2021年第4期299-305,共7页Journal of Jimei University：Natural Science

基　　金：福建省种业创新与产业化工程项目(2017FJSCZY03);福建省海洋渔业结构调整专项(2017HYJG07,2017HYJG08);2019年度福建省海洋经济发展专项;省属公益类科研院所专项(2020R1013008)。

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_(v)WD),并采用Biacore-表面等离子体共振(SPR)系统检测了rTF_(v)WD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_(v)WD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_(v)WD结合后,TTX的毒性降低了。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_(v)WD)of Takifugu flavidus in pichia pastoris was established and the target recombinant protein was successfully expressed in this study.The affinity between rTF_(v)WD and TTX was determined by using Biacore-surface plasma resonance(SPR)system.Intraperitoneal injection of mice was used to verify the neutralization of rTF_(v)WD and TTX in vivo.The results showed that the equilibrium dissociation constant(K D)of rTF_(v)WD and TTX was 3.1 mmol/L;When TTX and rTF_(v)WD 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.

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 

分 类 号：Q786[生物学—分子生物学]