Abstract
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 (Takeda et al., 2000. J. Virol. 74, 6643-6647), and the recovery rate was 100%. The wild-type MeV that encodes the lac-Z gene of approximately 3.2 kb 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.
| Original language | English |
|---|---|
| Pages (from-to) | 161-165 |
| Number of pages | 5 |
| Journal | Virus Research |
| Volume | 108 |
| Issue number | 1-2 |
| DOIs | |
| Publication status | Published - Mar 1 2005 |
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All Science Journal Classification (ASJC) codes
- Cancer Research
- Virology
- Infectious Diseases
Cite this
Efficient rescue of measles virus from cloned cDNA using SLAM-expressing Chinese hamster ovary cells. / Takeda, Makoto; Ohno, Shinji; Seki, Fumio; Hashimoto, Koji; Miyajima, Naoko; Takeuchi, Kaoru; Yanagi, Yusuke.
In: Virus Research, Vol. 108, No. 1-2, 01.03.2005, p. 161-165.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Efficient rescue of measles virus from cloned cDNA using SLAM-expressing Chinese hamster ovary cells
AU - Takeda, Makoto
AU - Ohno, Shinji
AU - Seki, Fumio
AU - Hashimoto, Koji
AU - Miyajima, Naoko
AU - Takeuchi, Kaoru
AU - Yanagi, Yusuke
PY - 2005/3/1
Y1 - 2005/3/1
N2 - 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 (Takeda et al., 2000. J. Virol. 74, 6643-6647), and the recovery rate was 100%. The wild-type MeV that encodes the lac-Z gene of approximately 3.2 kb 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.
AB - 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 (Takeda et al., 2000. J. Virol. 74, 6643-6647), and the recovery rate was 100%. The wild-type MeV that encodes the lac-Z gene of approximately 3.2 kb 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.
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UR - http://www.scopus.com/inward/citedby.url?scp=12844270574&partnerID=8YFLogxK
U2 - 10.1016/j.virusres.2004.09.002
DO - 10.1016/j.virusres.2004.09.002
M3 - Article
C2 - 15681066
AN - SCOPUS:12844270574
VL - 108
SP - 161
EP - 165
JO - Virus Research
JF - Virus Research
SN - 0168-1702
IS - 1-2
ER -