Heino P, Dillner J, Schwartz S
Microbiology and Tumorbiology Center, Karolinska Institute, Stockholm, Sweden.
Virology. 1995 Dec 20;214(2):349-59. doi: 10.1006/viro.1995.0044.
Several neutralizing sites of the human papillomavirus (HPV) capsid are known to be critically dependent on the conformation of the capsid. However, efficient production of HPV16 capsids in mammalian cells has been difficult, possibly because the HPV genome contains negative regulatory elements. To circumvent these problems, we cloned the HPV16 L1 and L2 genes from a healthy HPV16-infected woman into a Semliki Forest virus based expression vector (P. Liljeström and H. Garoff, Biotechnology 9, 1356-1361, 1991). Recombinant HPV16 L1- or L2-producing Semliki Forest virus was generated and used for infection of mammalian cells. The HPV16 L1 and L2 proteins were efficiently expressed and the majority of the L1 protein self assembled into virus-like particles (VLPs). Coexpression of L1 and L2 resulted in incorporation of L2 into the VLPs. The particles had a density of approximately 1.3 g/ml as determined by density gradient centrifugation. Transmission electron microscopy revealed that the particles had a morphology similar to native virions. The HPV16 VLPs produced by the Semliki Forest virus expression system may be useful as a conformationally correctly assembled target for studies of HPV attachment, assembly, serology, or vaccination.
已知人乳头瘤病毒（HPV）衣壳的几个中和位点严重依赖于衣壳的构象。然而，在哺乳动物细胞中高效生产HPV16衣壳一直很困难，这可能是因为HPV基因组包含负调控元件。为了规避这些问题，我们从一名健康的HPV16感染女性中克隆了HPV16的L1和L2基因，并将其插入到基于Semliki森林病毒的表达载体中（P. Liljeström和H. Garoff，《生物技术》9，1356 - 1361，1991）。产生了重组的表达HPV16 L1或L2的Semliki森林病毒，并用于感染哺乳动物细胞。HPV16的L1和L2蛋白得到了高效表达，并且大部分L1蛋白自组装成病毒样颗粒（VLPs）。L1和L2的共表达导致L2掺入到VLPs中。通过密度梯度离心测定，这些颗粒的密度约为1.3 g/ml。透射电子显微镜显示这些颗粒的形态与天然病毒粒子相似。由Semliki森林病毒表达系统产生的HPV16 VLPs可作为构象正确组装的靶标，用于HPV附着、组装、血清学或疫苗接种的研究。
