Suppression subtractive hybridization method for the identification of a new strain of murine hepatitis virus from xenografted SCID mice.
During attempts to clone retroviral determinants associated with a mouse model of Langerhans cell histiocytosis (LCH), suppression subtractive hybridization (SSH) was used to identify unique viruses in the liver of severe combined immunodeficiency (SCID) mice transplanted with LCH tissues. A partial genomic sequence of a murine coronavirus was identified, and the whole genome (31428 bp) of the coronavirus was subsequently sequenced using PCR cloning techniques. Nucleotide sequence comparisons revealed that the genome sequence of the new virus was 91-93% identical to those of known murine hepatitis viruses (MHVs). The predicted open reading frame from the nucleotide sequence encoded all known proteins of MHVs. Analysis at the protein level showed that the virus was closely related to the highly virulent MHV-JHM strain. The virus strain was named MHV-MI. No type D retroviruses were found. Degenerate PCR targeting of type D retrovirus and 5'-RACE targeting of other types of retroviruses confirmed the absence of any retroviral association with the LCH xenografted SCID mice.
Fiona Elsey Cancer Research Institute, 106-110 Lydiard Street South, Ballarat Technology Park Central, Ballarat, VIC, 3353, Australia. monirul@fecri.org.au.|School of Applied and Biomedical Sciences, Federation University Australia, Ballarat, VIC, 3350, Australia. monirul@fecri.org.au.
Mohammed M
MM
Islam
Fiona Elsey Cancer Research Institute, 106-110 Lydiard Street South, Ballarat Technology Park Central, Ballarat, VIC, 3353, Australia.
Brendan
B
Toohey
Department of Microbiology and Immunology, Peter Doherty Institute for Infection and Immunity, The University of Melbourne, Parkville, VIC, 3010, Australia.
Damian F J
DF
Purcell
Fiona Elsey Cancer Research Institute, 106-110 Lydiard Street South, Ballarat Technology Park Central, Ballarat, VIC, 3353, Australia.|School of Applied and Biomedical Sciences, Federation University Australia, Ballarat, VIC, 3350, Australia.
George
G
Kannourakis
D000293:Adolescent; D000818:Animals; D001483:Base Sequence; D018352:Coronavirus Infections / Q000821:virology*; D004195:Disease Models, Animal; D005260:Female; D016679:Genome, Viral; D006646:Histiocytosis, Langerhans-Cell / Q000821:virology*; D006801:Humans; D051379:Mice; D016513:Mice, SCID; D008969:Molecular Sequence Data; D006517:Murine hepatitis virus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction; D066251:Subtractive Hybridization Techniques / Q000379:methods*; D023041:Xenograft Model Antitumor Assays