【摘要】 目的 从感染HTLV-1的中国患者外周血单核细胞（PBMCs）中扩增编码HTLV-1核心蛋白P24的cDNA,并在大肠杆菌中进行表达。方法 提取感染者PBMCs基因组DNA,应用巢式PCR方法扩增出HTLV-1核心蛋白P24的cDNA并测序,与pGEX-20T载体构建重组质粒,在大肠杆菌BL21中表达,采用ELISA和Westernblot分析重组蛋白活性。结果HTLV-1核心蛋白P24基因序列高度保守,构建重组载体后,在大肠杆菌中表达的重组蛋白,经检测具有较强的抗原活性。结论 成功地表达了HTLV-1型病毒的核心蛋白P24基因,为国产诊断试剂和疫苗的研发打下了基础。更多还原

【Abstract】 Objective To amplify the cDNA encoding core protein P24 from the PBMCs of a Chinese patient infected with human T-cell lymphotropic virus type 1 ( HTLV - 1) and express in E. coli. Methods Extract DNA from the patient, amplify p24 gene by nested - PCR and sequence, then clone into vector pGEX -20T and express in E. coli. The expressed product was identified by Western blot and detected for activity by ELISA. Results The sequence of p24 gene was highly conserved, and the p24 protein with good antigenicity was successfully expressed in E. coli. Conclusion The cloning and successful expression of core protein p24 gene lays a foundation of developing diagnostic kit and vaccine.更多还原