In situ analysis of latency-associated transcripts as an indicator of latent infection of Marek's disease virus.

点击翻译按钮获取中文摘要

The purpose of this study is the transcriptional analysis of latently MDV-infected cells. In 1996 fiscal year, we made modification and became skillful in the methods for the analysis. Since low copies of MDV-trancripts would express in latently infected cells, we estimated reverse transcriptase-polymerase chain reaction (RT-PCR) , in situ hybridization and immunohistochemical techniques. In 1997 fiscal year, we detected newly identified latency-associated transcript (LAT) from MDV-transformed cells, which is known as a model of latent infection of MDV.In the MDV-transformed cells, spontaneously activated MDV is rarely included while it may influence the result of transcriptional analysis. We showed that a population consisting of 10^3 MDV-transformed cells probably did not include spontaneously activated MDV from immunohistochemical and RT-PCR-limiting-dilution analysis of MDV-transformed cells. In investigated genes (ICP4, ICP27, po1, TK,US3, A41, gA,gB,UL50, VP16andSORF2) a transcript corresponding to the ICP4 sequence was detected as a 0.7kbp RT-PCR product in 10^3 cells of MSB-1, RPL-1, HPRS-1, MOGA-1, MOGA-2, MSB-1 and MTB-1 but not in the retrovirus-transformed 1104B1 cell line. The transcript corresponding to the 0.7 kbp RT-PCR product suggested to be a common transcript in latently infected MDV-transformed cells, while most of the genes did not transcribe in these cells.