Murine gammaherpesvirus 68 (MHV-68) is a worldwide model which serves for the study of various aspects of gammaherpesviral infection. In 1976, the prototype strain and 4 isolates 72, 76, 78) were isolated from small rodents caught near Bratislava, Slovakia (1). Isolate MHV-Šumava was acquired few years later from Myodes glareolus caught in Šumava, Czech Republic (8). All isolates were passaged since then on various cell lines for the purpose of biological characterization and in vitro studies. Even though only MHV68´s genome is fully sequenced and therefore is the most intensively studied, a considerable amount of studies is focused also on characterization of other MHV isolates, particularly on their pathogenic, immunological and oncogenic properties in vivo. Besides variations during pathogenesis, MHV isolates remarkably diff ered also in their ability to cause lymphomas in the course of long-term infection. Th e most oncogenic isolate is MHV-60, which caused tumor development in 22% of chronically infected mice (10). MHV-76 was the only one from MHV isolates that did not show any signifi cant tumor development aft er 2-years long infection of mice (6). Since genomes of MHV isolates are not completely sequenced, it has to be determined, what is responsible for their diff erent pathogenic and oncogenic properties. In our laboratory, we have all above mentioned MHV isolates and we intended to make basic comparative analysis of their genomes in an eff ort to fi nd a genetic basis for their various pathogenic properties and oncogenic potential. Moreover, we tested two variants of MHV-76 -original isolate MHV-76 lyophilized in 1989 marked as MHV-76 (´89) and isolate MHV-76 (´08) which underwent multiple passages on diff erent cell lines until year 2008. Also, two variants of MHV-72 were used -MHV-72 (´97) stock frozen in 1997 and isolate MHV-72 (´08) propagated through multiple cell lines until 2008.First we analyzed MHV isolate´s DNA by RFLP method to identify larger portions of deletions or insertions and more remarkable diff erences in compared genomes to pinpoint the regions of interest in MHV genomes for further analysis. For RFLP analysis, viral DNA was extracted from virions present in supernatants of NIH 3T3 cells infected with MHV isolates at low multiplicity of infection. Viral DNA was digested overnight with EcoRI (Promega), BamHI (Promega) and HindIII (Th ermo Scientifi c). Digested DNA fragments were separated by electrophoresis, which was carried out in horizontal 0.7% agarose gel. Restriction profi le analysis of MHV-68 and MHV isolates showed that the most diff erences among isolates were found in fragments that represented the left end of the MHV genome -absence of fragment corresponding to nucleotides 1571-14270 (aft er 112 LETTERS TO THE EDITOR digestion with EcoRI) and 107-6262 (digestion with HindIII). Here are located genes responsible for regulation of pathogenic processes in vivo and oncogenesis not only in MHV but also in genomes of EBV and KSHV, and therefore we wanted to map this region...