Analysis of Kaposi's sarcoma-associated herpesvirus latent replication using a real-time polymerase chain reaction technique
Autor: | Taegun Seo, Yejin Kim, Jun Hyeong Jang, Lee Rang Hwang, Seho Cha |
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Rok vydání: | 2013 |
Předmět: |
viruses
virus diseases Nuclear Proteins Biology medicine.disease_cause Real-Time Polymerase Chain Reaction Virus Replication Virology law.invention Real-time polymerase chain reaction Viral replication Lytic cycle law Host chromosome DNA Viral Herpesvirus 8 Human medicine Vector (molecular biology) Kaposi's sarcoma-associated herpesvirus Antigens Viral Polymerase chain reaction Southern blot |
Zdroj: | Journal of virological methods. 193(2) |
ISSN: | 1879-0984 |
Popis: | Kaposi's sarcoma-associated herpesvirus (KSHV) undergoes replication independently via latent and lytic pathways. Latent replication is mediated by latent-associated nuclear antigen (LANA), the sole viral trans element for genome maintenance and replication. According to previous studies, LANA tethers the KSHV genome to the host chromosome during latency and interacts with host factors to ensure proper latent replication. Studies using Southern blot experiments have revealed consistently that vector constructs containing the viral terminal repeat (TR) region as a cis element in latent replication are replicated in the presence of LANA. However, Southern blotting is a time-intensive, complicated technique that requires multiple reagents. In addition, it has a limited ability to detect slight changes in replication efficiency under different conditions owing to its relatively low sensitivity. In the current study, a real-time polymerase chain reaction method was developed for detecting transient KSHV replication and was found to be capable of further identifying several factors that affect latent replication. This technique should provide a useful tool for the detection of KSHV latent replication under various conditions, including overexpression of viral or cellular factors and chemical stimulation. |
Databáze: | OpenAIRE |
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