国家自然科学基金（31302096）, 江苏省农业支撑项目（BE2013415）, 江苏省自然科学基金项目（BK2011536）, 江苏省”企业博士集聚计划”和江苏省”六大人才高峰项目”（NY-009）共同资助。
根据血清1型鸭甲型肝炎病毒(DHAV-1) SH株3D基因序列设计并合成1对特异性引物，通过RT-PCR方法扩增3D基因，将其克隆入原核表达载体pET-32a，转化大肠杆菌BL21(DE3)，在IPTG的诱导培养下重组蛋白获得了成功表达。SDS-PAGE显示重组蛋白的分子量约为68 kD，Western blot分析显示该重组蛋白能与多聚组氨酸标签单抗发生特异性反应。将切胶后纯化的目的蛋白免疫ICR小鼠，制备针对重组蛋白的多抗血清，Western blot结果显示制备的抗血清能够与DHAV-1感染的SPF鸡胚尿囊液总蛋白发生特异反应。以上结果表明，3D基因在大肠杆菌中获得了成功表达，且制备的多抗血清可以用于3D蛋白的检测，为3D蛋白的功能研究奠定了基础。
According to the genome sequences of duck hepatitis A virus type-1(DHAV-1), one pair of specific primers was designed to amplify the 3D gene using RT-PCR. The amplified fragment was cloned into prokaryotic expression vector pET-32a and the recombinant plasmid was transformed into BL21(DE3) E.coli. Recombinant proteins were successfully expressed following IPTG induction. SDS-PAGE showed that the recombinant expression protein was about 68 kD. Western blot assay revealed that the desired proteins could be recognized by the monoclonal antibody against histidine-tagged proteins. The antiserum against the recombinant protein was produced by the immunized ICR mouse with recombinant proteins. Western blot results showed that the antiserum could react specifically with DHAV-1 allantoic fluid. These results showed that the 3D gene was successfully expressed in E.coli and the antiserum could be used for detection of the 3D protein, which would lay a foundation for function study of the 3D protein.