CAJ | 학술논문

为探索鸭肠炎病毒（ DEV）囊膜糖蛋白gE 基因的相关特性和功能，应用PCR扩增了DEV的gE基因，全长1473 bp，编码490个氨基酸，其中1～22位氨基酸为信号肽。 gE基因与国内分离强毒CHv株、国内疫苗株同源性均为100％，与德国分离强毒2085株同源性达到99％，显示其高度保守。将gE基因完整开放阅读框（ gE）和去信号肽的gE基因（ gE’）分别克隆至真核表达载体pCDNA3．1，获得重组质粒 pCDNA3．1-gE、pCDNA3．1-gE，然后应用脂质体法转染 vero 细胞。RT-PCR扩增证实gE、gE’均在细胞内进行了转录，而间接免疫荧光试验证实只有gE’基因在vero细胞中获得了表达，为研究gE蛋白的功能及研制鸭瘟抗体检测试剂盒奠定了基础。
위탐색압장염병독（ DEV）낭막당단백gE 기인적상관특성화공능，응용PCR확증료DEV적gE기인，전장1473 bp，편마490개안기산，기중1～22위안기산위신호태。 gE기인여국내분리강독CHv주、국내역묘주동원성균위100％，여덕국분리강독2085주동원성체도99％，현시기고도보수。장gE기인완정개방열독광（ gE）화거신호태적gE기인（ gE’）분별극륭지진핵표체재체pCDNA3．1，획득중조질립 pCDNA3．1-gE、pCDNA3．1-gE，연후응용지질체법전염 vero 세포。RT-PCR확증증실gE、gE’균재세포내진행료전록，이간접면역형광시험증실지유gE’기인재vero세포중획득료표체，위연구gE단백적공능급연제압온항체검측시제합전정료기출。
To explore the correlated properties and functions of glycoprotein gE in duck enteritis virus( DEV) ,the sequence of gE was amplified by PCR in this study. The whole length of gE open reading frame( ORF) is 1473 bp and encodes 490 amino acids, 1to 22 amino acids of which were defined as signal peptide. Gene gE of DEV strain AV1221 share 100% homology with gE of both virulent strain and vaccine strain in China,also 99%with virulent strain 2085 in German, which shows gE of DEV strain AV1221 is highly homological. The complete ORF ( gE) and truncated glycoprotein gE without signal peptide ( gE’ ) were respectively cloned to the eukaryotic expression vetor pCDNA3.1. Recombinant pCDNA3.1-gE and pCDNA3.1-gE’ plasmids were transfected to vero cells with liposome mediated transfection method. Successful transcription of gE and gE ’ in vero cells were confirmed by RT-PCR, while successful expression of gE’ alone in vero cells was verified by indirect immunofluorescence assay. The study provides a fundation for function of DEV gE and development of gE antibody test kit.