马立克氏病病毒 RB1B 株 UL14蛋白原核表达及多抗血清制备

摘要

为研究 UL14蛋白在马立克氏病病毒感染过程中的生物学作用，本文以 RB1B 毒株感染的鸡胚成纤维细胞基因组 DNA为模板，经 PCR 扩增出 UL14基因，然后克隆入 pET32a 载体，转化 BL21（DE3）工程细菌且经 IPTG 诱导进行原核表达。结果显示，pET32a-UL14重组质粒在大肠杆菌中表达，表达的重组蛋白具有可溶性。IFA 分析证实，用此融合蛋白免疫 BALB/c小鼠后获得的多抗血清能与 MDV RB1B 感染的 CEF 细胞发生特异性反应。本研究结果为 UL14蛋白的生物学功能研究提供了条件。%To investigate the biological function of UL14 protein during the course of MDV infection, UL14 gene from CEF cells infected with RB1B strain was amplified by PCR. UL14 gene was cloned into pET32a (+) vector and expressed in E.coli BL21 (DE3). Recombinant His-UL14 protein expressed in E.coli was soluble. The BALB/c mice were immunized with recombinant UL14 protein and resulting antiserum was used in IFA. The specific inflorescence was visualized in CEF cells infected with MDV strain RB1B .