Molecular cloning and expression of a spike protein of neurovirulent murine coronavirus JHMV variant cl-2.
A cDNA encoding the spike (S) protein of the neurovirulent murine coronavirus JHMV variant cl-2 was isolated and sequenced. Analysis of the cDNA revealed that the S protein consists of 1376 amino acids, as does the S protein of mouse hepatitis virus 4. We inserted the cDNA into the genome of vaccinia virus to obtain a recombinant vaccinia virus (rVV). The S protein expressed in RK13 cells infected by the rVV was shown to be electrophoretically and immunologically indistinguishable from the S protein produced in DBT cells infected with cl-2 virus. RVV infection of rats and mice induced S protein-specific antibody production detectable by immunofluorescence and neutralization. Moreover, the S protein expressed by the rVV induced syncytium formation not only in mouse DBT and L cells, which are susceptible to cl-2 virus infection, but also in rabbit RK13 cells, which are not susceptible to cl-2 virus infection. This result suggests the possibility that RK13 cells have binding sites for the cl-2 virus S protein.
Taguchi
F
F
National Institute of Neuroscience, Tokyo, Japan.
Ikeda
T
T
Shida
H
H
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D003001:Cloning, Molecular; D015726:Giant Cells; D006023:Glycoproteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D008562:Membrane Glycoproteins; D051379:Mice; D008969:Molecular Sequence Data; D006517:Murine hepatitis virus / Q000235:genetics* / Q000276:immunology / Q000472:pathogenicity; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology
D000911:Antibodies, Monoclonal; D006023:Glycoproteins; D008562:Membrane Glycoproteins; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins; C578557:spike glycoprotein, SARS-CoV; C578553:spike protein, mouse hepatitis virus