Submitted to: Differential Display and Related Techniques for Gene Discovery
Publication Type: Abstract Only
Publication Acceptance Date: October 6, 1996
Publication Date: N/A
Technical Abstract: Marek's disease virus (MDV) is an avian herpesvirus which causes lymphoproliferative disease in chicken. Attenuation of MDV by passage in tissue culture is well established. During the process of the attenuation, several changes in the virus genome have been reported. However, it is not clear which change(s) on the genome is responsible for the attenuation. We have applied differential display technique to the RNA from virulent and passage-attenuated MDV-infected chicken embryo fibroblast in order to detect differences in mRNA expression in these cells. We have examined 104 pairs of primers (26 5' primers x 4 3' primers) and 116 specific bands in virulent MDV-infected cells could be recovered after reamplification. Among them, we found four species of fragments which have homology to the MDV genome. Three were amplified with 5' and 3' primers. One corresponds to the 3' end of reported VP16 mRNA. The second fragment was amplified from an internal poly(T) stretch with the 3' primer and corresponding to reported sequence located at 5' to ICP4 coding region. The third fragment contained two short stretches of MDV sequences separated by an unknown sequence, and was amplified from with 5' and 3' primers. The fourth fragment was amplified with two 5' primers and has a sequence around the virus genome region where changes during the attenuation have been reported. In Northern blots, all fragments, except the third fragment, hybridized to multiply transcripts in both virulent and attenuated virus-infected cells. No quantitative difference in expression levels could be detected. However, the fourth fragment detected a transcript which seemed to be different in size between these viruses.