本文選題：呼吸道合胞病毒 + 融合糖蛋白��； 參考：《北京交通大學(xué)學(xué)報(bào)》2016年06期

【摘要】：分析以人呼吸道合胞病毒(Human respiratory syncytial virus,RSV)融合糖蛋白(fusion glycoprotein,F)為抗原靶向DEC205/CD205受體策略對(duì)重組F蛋白表達(dá)及活性的影響.F蛋白是RSV的重要中和抗原,克隆RSV F蛋白胞外區(qū)的編碼基因至可識(shí)別鼠樹(shù)突狀細(xì)胞(Dendritic cells,DCs)DEC205受體的單鏈抗體(single chain antibody of anti-DEC205,scDEC)的C端,構(gòu)建可表達(dá)重組F蛋白(scDECF)的質(zhì)粒pVAX1/scDECF,Western blot方法檢測(cè)scDECF的表達(dá),免疫熒光法和流式細(xì)胞術(shù)檢測(cè)表達(dá)的scDECF與DEC205受體的結(jié)合活性.經(jīng)Western blot證實(shí)在pVAX1/scDECF轉(zhuǎn)染的293T細(xì)胞成功表達(dá)了scDECF.將所獲得的細(xì)胞培養(yǎng)上清與表達(dá)鼠DEC205受體的細(xì)胞CHOmDEC205孵育,用免疫熒光和流式細(xì)胞術(shù)證實(shí):與作為對(duì)照的重組F蛋白scISOF相比,所獲得的scDECF與CHOmDEC205細(xì)胞呈現(xiàn)明顯的結(jié)合.本研究成功表達(dá)了重組蛋白scDECF,且所獲得的重組蛋白scDECF具有與表達(dá)DEC205受體的細(xì)胞特異性結(jié)合的活性.
[Abstract]:To analyze the effect of fusion glycoprotein (DEC205/CD205) of human respiratory syncytial virus (respiratory syncytial virus) on the expression and activity of recombinant F protein. F protein is an important neutralizing antigen of RSV. The encoding gene of extracellular domain of RSV F protein was cloned to the C-terminal of single chain antibody of anti-DEC205 (single chain antibody of anti-DEC205), which could recognize the receptor of mouse dendritic cells. The plasmid pVAX1 / scDECFWestern blot was used to detect the expression of scDECF. The binding activity of scDECF to DEC205 receptor was detected by immunofluorescence and flow cytometry. The expression of scDECF in 293T cells transfected with pVAX1/scDECF was confirmed by Western blot. The obtained supernatants were incubated with CHOmDEC205 cells expressing mouse DEC205 receptor. Immunofluorescence and flow cytometry showed that the scDECF was significantly bound to CHOmDEC205 cells compared with the recombinant F protein scISOF as control. In this study, the recombinant protein scDECF was successfully expressed, and the recombinant protein scDECF had specific binding activity to the cells expressing DEC205 receptor.
【作者單位】： 北京交通大學(xué)生命科學(xué)與生物工程研究院;
【基金】：中央高�；究蒲袠I(yè)務(wù)費(fèi)專項(xiàng)資金資助(2009YJS037) 國(guó)家重大新藥創(chuàng)制專項(xiàng)資金資助(2013ZX09103-003-011)
【分類號(hào)】：R373.14

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