目的  采用昆虫细胞-杆状病毒表达系统扩增人乳头瘤病毒（human papillomavirus，HPV）58 型 L1 蛋白基因，确定 L1 蛋白表达的最佳条件。 方法   取处于对数生长期的Sf 9 细胞以不同的感染复数（MOI）值扩增病毒，用噬斑试验检测病毒滴度，确定扩增重组病毒的最适条件；用间接免疫荧光法判断目的蛋白表达情况；通过蛋白印迹法测定不同的表达时间和 MOI 值条件下目的蛋白表达量，确定目的蛋白的最佳表达条件。 结果  处于对数生长期的细胞以 MOI 值为 0.5 表达48 h 条件下扩增含 HPV58 L1 基因的重组杆状病毒，滴度最高，可达 5×108 pfu∕ml；以 MOI 值为 5.0 表达 72 h 获得最佳的目的蛋白表达量。 结论  确定了扩增病毒和表达相应目的蛋白的最佳条件。

Objective  To select the optimum conditions for amplification and expression of the recombinant human papillomavirus 58(HPV58) L1 gene using baculovirus-insect cell expression system.  Methods  The Sf 9 insect cells at logarithmic phase was used to amplify virus at different multiplicities of infection (MOI). Titers of the virus were determined by plaque assay. The expression of recombinant protein was identified by indirect immunofluorescence assay, and the expression of recombinant protein at different MOI and time was determined by Western blot.  Results  Recombinant baculovirus containing HPV58 L1 gene were amplified at MOI of 0.5 for 48 h when the Sf 9 insect cells were at logarithmic phase, and the highest titers of the virus reached 5×108 pfu/ml. The optimum conditions for expressing the recombinant protein were that the recombinant baculovirus was inoculated at MOI of 5.0 and the protein was expressed within 72 h. Conclusion The optimum conditions for amplifying the virus and expressing the recombinant protein are determined.