中国动物传染病学报
中國動物傳染病學報
중국동물전염병학보
CHINESE JOURNAL OF VETERINARY PARASITOLOGY
2014年
3期
19-25
,共7页
黎摄儿%赵苗苗%崔成%黎满香
黎攝兒%趙苗苗%崔成%黎滿香
려섭인%조묘묘%최성%려만향
羊传染性脓疱病毒%毒力因子趋化因子结合蛋白%二级结构%细胞表位%预测
羊傳染性膿皰病毒%毒力因子趨化因子結閤蛋白%二級結構%細胞錶位%預測
양전염성농포병독%독력인자추화인자결합단백%이급결구%세포표위%예측
Orf virus%chemokine-binding protein%secondary structure%epitope%prediction
参照GenBank中羊传染性脓疱病毒（Orf virus, ORFV）的毒力因子趋化因子结合蛋白（chemokine-binding protein， CBP）基因的核苷酸序列设计合成1对特异性引物，以羊传染性脓疱病毒湖北分离毒株的DNA为模板，提取总DNA。采用PCR方法特异扩增该基因片段，得到CBP基因序列。通过网络平台的SOPMA服务器和DNAStar软件预测ORFV CBP蛋白的二级结构；综合利用DNAStar软件、二级结构预测及ABCpred方案预测羊ORFV CBP蛋白的B细胞表位；分别采用人工神经网络法（ANNA）和在线程序预测ORFV CBP蛋白的细胞毒性T细胞和辅助T细胞的抗原表位。结果显示，ORFV CBP蛋白的二级结构主要包括α-螺旋、β-转角、无规则卷曲和β-片层4种类型，分别为α-螺旋占24.64％、β-片层占22.14％、β-转角占3.39％、无规则卷曲占49.29％；有7个优势B细胞表位，7个细胞毒性T细胞表位和3个辅助T细胞表位。本研究为ORFV诊断方法的建立、单克隆抗体的制备及表位疫苗的研制提供了理论依据。
參照GenBank中羊傳染性膿皰病毒（Orf virus, ORFV）的毒力因子趨化因子結閤蛋白（chemokine-binding protein， CBP）基因的覈苷痠序列設計閤成1對特異性引物，以羊傳染性膿皰病毒湖北分離毒株的DNA為模闆，提取總DNA。採用PCR方法特異擴增該基因片段，得到CBP基因序列。通過網絡平檯的SOPMA服務器和DNAStar軟件預測ORFV CBP蛋白的二級結構；綜閤利用DNAStar軟件、二級結構預測及ABCpred方案預測羊ORFV CBP蛋白的B細胞錶位；分彆採用人工神經網絡法（ANNA）和在線程序預測ORFV CBP蛋白的細胞毒性T細胞和輔助T細胞的抗原錶位。結果顯示，ORFV CBP蛋白的二級結構主要包括α-螺鏇、β-轉角、無規則捲麯和β-片層4種類型，分彆為α-螺鏇佔24.64％、β-片層佔22.14％、β-轉角佔3.39％、無規則捲麯佔49.29％；有7箇優勢B細胞錶位，7箇細胞毒性T細胞錶位和3箇輔助T細胞錶位。本研究為ORFV診斷方法的建立、單剋隆抗體的製備及錶位疫苗的研製提供瞭理論依據。
삼조GenBank중양전염성농포병독（Orf virus, ORFV）적독력인자추화인자결합단백（chemokine-binding protein， CBP）기인적핵감산서렬설계합성1대특이성인물，이양전염성농포병독호북분리독주적DNA위모판，제취총DNA。채용PCR방법특이확증해기인편단，득도CBP기인서렬。통과망락평태적SOPMA복무기화DNAStar연건예측ORFV CBP단백적이급결구；종합이용DNAStar연건、이급결구예측급ABCpred방안예측양ORFV CBP단백적B세포표위；분별채용인공신경망락법（ANNA）화재선정서예측ORFV CBP단백적세포독성T세포화보조T세포적항원표위。결과현시，ORFV CBP단백적이급결구주요포괄α-라선、β-전각、무규칙권곡화β-편층4충류형，분별위α-라선점24.64％、β-편층점22.14％、β-전각점3.39％、무규칙권곡점49.29％；유7개우세B세포표위，7개세포독성T세포표위화3개보조T세포표위。본연구위ORFV진단방법적건립、단극륭항체적제비급표위역묘적연제제공료이론의거。
According to the published chemokine-binding protein (CBP) gene sequence of Orf virus in the GenBank, a pair of primers were designed and synthesized for amplification in PCR using total DNA of Orf virus Hubei strain as template. The PCR products of the CBP gene were sequenced. Subsequently, the secondary structure of the CBP protein was predicted using the network platform SOPMA server and DNAStar software. The B cell epitopes of the CBP protein were predicted by comprehensive utilization of the DNAStar software, the secondary structure prediction and ABCpred program. The cytotoxic T cell and T helper epitopes of the CBP protein were predicted using the artificial neural network (ANNA) and online program. The results showed that the CBP protein of Orf virus was composed withα-helixes (24.64%),β-sheet (22.14%), random coils (49.29%) andβ-turn (3.39%). Prediction of epitomes revealed that the CBP protein had 7 B cell epitopes, 7 cytotoxic T cell epitopes and 3 T helper epitopes. The information obtained from these predictions provided theoretical foundations for development of diagnostic methods and vaccines as well as preparation of monoclonal antibodies against Orf virus.
