人乳头瘤病毒类病毒的产生及其对原代人角质形成细胞的感染。
Human Papillomavirus Quasivirus Production and Infection of Primary Human Keratinocytes.
机构信息
Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institues of Health, Bethesda, Maryland.
Biological Science Graduate Program, University of Maryland, College Park, Maryland.
出版信息
Curr Protoc Microbiol. 2020 Jun;57(1):e101. doi: 10.1002/cpmc.101.
This protocol describes the production of human papillomavirus (HPV)-derived quasiviruses. Quasiviruses are infectious particles that are produced in 293TT packaging cells and contain a complete viral genome. We describe methods for infection of primary human keratinocytes with HPV quasiviruses, as well as assays to measure early viral DNA replication and transcription. Published 2020. U.S. Government. Basic Protocol 1: Transfection, harvest, and isolation of HPV quasiviruses Alternate Protocol 1: Packaging HPV DNA replicated in 293TT cells Alternate Protocol 2: Production of higher-purity quasivirus using the "Ripcord" method Support Protocol 1: Production of HPV minicircles Support Protocol 2: Production of recircularized HPV genomes Support Protocol 3: Screening of fractions for viral proteins Support Protocol 4: Screening of fractions for viral DNA Support Protocol 5: Measuring viral titer Support Protocol 6: Quantitation of quasivirions Basic Protocol 2: Infection of primary human foreskin keratinocytes with quasivirus Basic Protocol 3: HPV quasivirus transcription assay Basic Protocol 4: HPV quasivirus replication assay.
本方案描述了人乳头瘤病毒（HPV）衍生类病毒的生产。类病毒是在 293TT 包装细胞中产生的传染性颗粒，包含完整的病毒基因组。我们描述了用 HPV 类病毒感染原代人角质形成细胞的方法，以及测量早期病毒 DNA 复制和转录的检测方法。 发表于 2020 年。美国政府。 基本方案 1：HPV 类病毒的转染、收获和分离 可选方案 1：在 293TT 细胞中复制的 HPV DNA 的包装 可选方案 2：使用“Ripcord”方法生产更高纯度的类病毒 支持方案 1：HPV 迷你环的生产 支持方案 2：回收 HPV 基因组的生产 支持方案 3：对病毒蛋白进行级分筛选 支持方案 4：对病毒 DNA 进行级分筛选 支持方案 5：测定病毒滴度 支持方案 6：类病毒粒子的定量 基本方案 2：用类病毒感染原代人包皮角质形成细胞 基本方案 3：HPV 类病毒转录分析 基本方案 4：HPV 类病毒复制分析。
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