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Journal of Virology, June 1999, p. 4919-4924, Vol. 73, No. 6
Department of Molecular Microbiology and
Immunology, Johns Hopkins School of Public Health, Baltimore,
Maryland 21205
Received 22 December 1998/Accepted 16 March 1999
The alphavirus Sindbis virus (SV) has a wide host range and infects
many types of cultured cells in vitro. The outcome of infection is
dependent on the strain of virus used for infection and the properties
of the cells infected. To identify cellular determinants of
susceptibility to SV infection we mutagenized Chinese hamster ovary
(CHO) cells by retroviral insertion with a vector containing the
neomycin resistance gene that allowed selection for integration into
transcriptionally active genes. Cells were then selected for survival
after infection with SV. The most resistant cell line (CHO-18.4m)
exhibited delayed virus replication and virus-induced cell death, had a
single retroviral insertion, and was defective in SV binding to the
cell surface. Further analysis revealed that CHO-18.4m cells were
deficient in the expression of the sulfated glycosaminoglycans heparan
sulfate and chondroitin sulfate. This further confirms the importance of heparan sulfate as an attachment molecule for SV in vitro and demonstrates the usefulness of this technique for identifying cellular
genes that are important for virus replication.
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Characterization of a Chinese Hamster Ovary Cell Line
Developed by Retroviral Insertional Mutagenesis That Is Resistant
to Sindbis Virus Infection

*
Corresponding author. Mailing address: Department of
Molecular Microbiology and Immunology, Johns Hopkins School of
Public Health, 615 N. Wolfe St., Rm. E5132, Baltimore, MD 21205. Phone: (410) 955-3459. Fax: (410) 955-0105. E-mail:
dgriffin{at}jhsph.edu.
Present address: National Defense Medical Center, Institute of
Preventive Medicine, Taipei, Taiwan, Republic of China.
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