PCR Detection of Murine Retroviruses and Characterization of a New Xenotropic Retrovirus from Myeloma Cells

Summary

We have established a ‘nested PCR’ technique for the group-specific detection of murine C-type retroviruses to screen cell lines for the presence of contaminants. Analyses confirmed that a well known myeloma cell line, Sp2/0, is contaminated with a retrovirus. The genome of the retrovirus was isolated and characterized by nucleic acid sequencing. The genomic organization of this retrovirus is typical for a murine C-type retrovirus. However, numerous differences could be found in the primary structure of gag and the integrase protein. The amino acid sequence of the envelope region and infection experiments with cell lines of different species indicate a xenotropic host range of the virus. The envelope region of the Sp2/0 virus shows strong homology to the xenotropic retrovirus found in CWD mice.