Acta Scientiarum Naturalium Universitatis Pekinensis

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In Vitro Construction of Pseudovirions of Human Papillomavirus Type 16

QIN Yan, CUI Honglian, HU Meihao   

  1. College of life sciences, Peking University, Beijing, 100871
  • Received:2005-06-17 Online:2006-03-20 Published:2006-03-20

人16型乳头瘤病毒“假病毒”的制备

秦焱,崔红莲,胡美浩   

  1. 北京大学生命科学学院,北京,100871

Abstract: The L1 major capsid protein of human papillomavirus type 16 was expressed in sf9 insect cells by recombinant baculoviruses and self-assembled into virus-like particles (VLPs). The VLPs were harvested by CsCl ultracentrifugation and were disassembled with DTT and EDTA. A reassembled process was performed by removing DTT and EDTA in the presence of E6E7-pcDNA3.1+ plasmid and Ca2+. The novel VLPs, in which the plasmid was packed, were named HPV16 pseudovirus.

Key words: human papillomavirus type 16, baculovirus expression system, pseudovirus

摘要: 利用杆状病毒表达体系,在 sf9 细胞中表达人16型乳头瘤病毒(Human Papillomavirus type 16, HPV16)的晚期表达蛋白L1,通过超速离心的方法得到HPV16病毒样颗粒(virus like particles, VLPs)。VLPs经过DTT和EDTA处理解聚成L1蛋白,将构建好的真核表达载体E6E7-pcDNA3.1+与解聚的L1蛋白混合,透析除去DTT和EDTA并逐渐升高Ca2+的浓度,解聚的L1能够重新聚集成VLPs,其中一部分VLPs包裹了真核表达载体,和天然的人乳头瘤病毒的构成很相似,作者称之为“假病毒”。

关键词: 人16型乳头瘤病毒, 杆状病毒表达系统, 假病毒

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