Human DNA contains sequences homologous to the 5'-non-coding region of hepatits C virus: characterization with restriction endonucleases reveals individual varieties.
OBJECTIVE
To investigate a 272 base pair section of the 5'-non-coding region of genomic DNA from the peripheral blood monounuclear cells of healthy hepatitis virus C (HCV)-negative human subjects (not patients).


METHODS
This sequence section bears interest because (1) it harbors several potential methylation (Cp-rich) sites, and (2) it represents the largest part of its internal ribosomal entry site. A pre-PCR digestion protocol was established making consistent use of four restriction endonucleases selected for certain features: SmaI, XmaCI, MspI, and HpaII are inhibited if methylation(s) are present at certain cytosines within their cutting sequences.


RESULTS
The suspected HCV-specific sequence was found in the DNA of each subject tested. The pre-PCR digestion assay reveals individual differences in their pattern of methylation, which may be due to possible epigenetic phenomena.


CONCLUSIONS
The results provide formal proof that these HCV-specific sequences are contained in the genomic or extra chromosomal target DNA, and probably belong to a new class of endogenous sequences.
Dennin
Reinhard H
RH
Institut für Medizinische Mikrobiologie und Hygiene, Universität zu Lübeck, Lübeck, Germany. dennin@hygiene.mu-luebeck.de
Wo
Jianer
J
D019175:DNA Methylation; D016174:Hepacivirus / Q000235:genetics*; D006801:Humans; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid