Avian Disease Research Center, College of Veterinary Medicine of Sichuan Agricultural University, Ya'an 625014, China.
Intervirology. 2009;52(6):326-34. doi: 10.1159/000242354. Epub 2009 Sep 29.
The UL24 gene of duck enteritis virus (DEV) is conserved across herpesviruses, but its protein characterization has not been reported. We expressed the UL24 gene in Escherichia coli BL21 from a recombinant plasmid pET32a/DEV-UL24 and used the resulting protein to raise antiserum. This antiserum recognized a 38-kDa protein in lysates from infected cells. SDS-PAGE analysis showed that the UL24 partial protein was highly expressed after induction by 0.4 mM IPTG at 30 degrees for 6 h. The results of purification revealed that expression protein was more purified using the method of electrophoresis than that of chromatography, but the yield was lower. In immunogenicity analysis, the protein could significantly elicit a specific antibody response in immunized ducklings when compared with the control groups, and the titers against expression protein reached the peak 1:5,120 (OD(450nm) = 2.5) on day 28 after immunization, while with mean titers of 1:10,240 (OD(450nm) = 3.37) in DEV commercial attenuated vaccine strain immunized duckling groups. It showed that expression protein is immunogenic in laboratory ducklings. On the basis of subcellular location, UL24 appeared to be predominantly nuclear membrane-associated, especially at later times in infection, and provided a good tool to further study the biofunctions of UL24 protein.
鸭肝炎病毒（DEV）的 UL24 基因在疱疹病毒中是保守的，但它的蛋白质特性尚未被报道。我们从重组质粒 pET32a/DEV-UL24 在大肠杆菌 BL21 中表达了 UL24 基因，并使用所得的蛋白质制备了抗血清。该抗血清在感染细胞的裂解物中识别出一种 38kDa 的蛋白质。SDS-PAGE 分析表明，在 30°C 下用 0.4mM IPTG 诱导 6 小时后，UL24 部分蛋白高度表达。纯化结果表明，与色谱法相比，电泳法更能纯化表达蛋白，但产量较低。在免疫原性分析中，与对照组相比，该蛋白在免疫鸭中能显著引起特异性抗体反应，免疫后第 28 天，表达蛋白的效价达到峰值 1:5,120（OD(450nm) = 2.5），而在 DEV 商业减毒疫苗株免疫鸭组中的平均效价为 1:10,240（OD(450nm) = 3.37）。这表明表达蛋白在实验室鸭中具有免疫原性。基于亚细胞定位，UL24 似乎主要与核膜相关，特别是在感染后期，为进一步研究 UL24 蛋白的生物功能提供了良好的工具。
