Suppression subtractive hybridization method for the identification of a new strain of murine hepatitis virus from xenografted SCID mice

Archives of Virology
Mohammed M IslamG Kannourakis

Abstract

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.

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Citations

Apr 17, 2021·Microbiology Resource Announcements·Amanda Barbosa GarciaClarice Weis Arns

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Datasets Mentioned

BETA
FJ647224.1
AY700211.1
AB551247

Methods Mentioned

BETA
PCR
xenografting
biopsy
electrophoresis

Software Mentioned

MegaBLAST
pDRAW32
Aca Clone
BLAST
pDRAW32 DNA Analysis
ClustalW2
GeneFisher2 Server
CLUSTAL W

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