Efficient rescue of measles virus from cloned cDNA using SLAM-expressing Chinese hamster ovary cells.
We here report a highly efficient reverse genetics system for measles virus (MeV), using Chinese hamster ovary cells constitutively expressing a MeV receptor human signaling lymphocyte activation molecule (CHO/hSLAM cells). The recombinant vaccinia virus vTF7-3 that encodes the T7 RNA polymerase under the control of the early/late promoter was used in the system. Replication of vTF7-3 was highly restricted in CHO/hSLAM cells, but the virus could still drive the T7 promoter, allowing us to recover MeV from the transfected cDNA efficiently. With this system the number of infectious centers, in which MeV replication cycles are initiated from transfected cDNAs, was approximately 100 times higher than that with the previous system (. J. Virol. 74, 6643-6647), and the recovery rate was 100%. The wild-type MeV that encodes the lac-Z gene of approximately 3.2kb in length, was easily generated with this CHO/hSLAM system, while such virus could not be recovered with the previous system. Since SLAM acts as a cellular receptor for both MeV vaccine and wild-type strains, the Edmonston vaccine strain was also recovered with this system more efficiently than with any other systems reported previously. Thus, the CHO/hSLAM-based system would expand applications of the MeV reverse genetics by allowing productions of mutant MeVs that have been difficult to generate with less efficient systems.
Takeda
Makoto
M
Department of Virology, Faculty of Medicine, Kyushu University, Fukuoka 812-8582, Japan. mtakeda@virology.med.kyushu-u.ac.jp
Ohno
Shinji
S
Seki
Fumio
F
Hashimoto
Koji
K
Miyajima
Naoko
N
Takeuchi
Kaoru
K
Yanagi
Yusuke
Y
D000818:Animals; D015703:Antigens, CD; D016466:CHO Cells; D002472:Cell Transformation, Viral; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary; D004279:DNA, Viral / Q000235:genetics / Q000502:physiology*; D017930:Genes, Reporter; D006023:Glycoproteins / Q000096:biosynthesis* / Q000235:genetics; D007136:Immunoglobulins / Q000096:biosynthesis* / Q000235:genetics; D008459:Measles virus / Q000235:genetics* / Q000502:physiology*; D011956:Receptors, Cell Surface; D011991:Receptors, Virus / Q000096:biosynthesis / Q000235:genetics; D000071177:Signaling Lymphocytic Activation Molecule Family Member 1; D014162:Transfection; D010948:Viral Plaque Assay; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D015703:Antigens, CD; D018076:DNA, Complementary; D004279:DNA, Viral; D006023:Glycoproteins; D007136:Immunoglobulins; D011956:Receptors, Cell Surface; D011991:Receptors, Virus; D000071177:Signaling Lymphocytic Activation Molecule Family Member 1; D001616:beta-Galactosidase