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A hybrid baculovirus-T7 RNA polymerase system for recovery of an infectious virus from cDNA.
Virology. 1997 May 12; 231(2):192-200.V

Abstract

We established a hybrid baculovirus-T7 RNA polymerase system for transient expression in mammalian cells. Two recombinant baculoviruses carrying cDNA of bacteriophage T7 RNA polymerase, with or without a nuclear localization signal, under the control of a mammalian promoter were constructed. High level expression of T7 RNA polymerase was observed in various mammalian cell lines after infection with the recombinant baculoviruses. After transfection of plasmids containing the luciferase gene under the control of the T7 promoter, high luciferase activity was detected in cells infected with the recombinant baculoviruses. We also constructed a plasmid containing an entire cDNA clone of type 1 poliovirus under the T7 promoter. Two days after transfection of the plasmid into the cells infected with the recombinant baculoviruses, a high titer of poliovirus was recovered. The use of the recombinant baculoviruses did not cause any cytopathic effects even at a high multiplicity of infection. The lack of replication ability and low toxicity are the advantageous features of the hybrid baculovirus-T7 polymerase system in comparison with the widely used vaccinia-T7 polymerase system for gene expression and recovery of infectious viruses from its cDNA.

Authors+Show Affiliations

Department of Virology II, National Institute of Health, Tokyo, Japan.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

9168881

Citation

Yap, C C., et al. "A Hybrid baculovirus-T7 RNA Polymerase System for Recovery of an Infectious Virus From CDNA." Virology, vol. 231, no. 2, 1997, pp. 192-200.
Yap CC, Ishii K, Aoki Y, et al. A hybrid baculovirus-T7 RNA polymerase system for recovery of an infectious virus from cDNA. Virology. 1997;231(2):192-200.
Yap, C. C., Ishii, K., Aoki, Y., Aizaki, H., Tani, H., Shimizu, H., Ueno, Y., Miyamura, T., & Matsuura, Y. (1997). A hybrid baculovirus-T7 RNA polymerase system for recovery of an infectious virus from cDNA. Virology, 231(2), 192-200.
Yap CC, et al. A Hybrid baculovirus-T7 RNA Polymerase System for Recovery of an Infectious Virus From CDNA. Virology. 1997 May 12;231(2):192-200. PubMed PMID: 9168881.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A hybrid baculovirus-T7 RNA polymerase system for recovery of an infectious virus from cDNA. AU - Yap,C C, AU - Ishii,K, AU - Aoki,Y, AU - Aizaki,H, AU - Tani,H, AU - Shimizu,H, AU - Ueno,Y, AU - Miyamura,T, AU - Matsuura,Y, PY - 1997/5/12/pubmed PY - 1997/5/12/medline PY - 1997/5/12/entrez SP - 192 EP - 200 JF - Virology JO - Virology VL - 231 IS - 2 N2 - We established a hybrid baculovirus-T7 RNA polymerase system for transient expression in mammalian cells. Two recombinant baculoviruses carrying cDNA of bacteriophage T7 RNA polymerase, with or without a nuclear localization signal, under the control of a mammalian promoter were constructed. High level expression of T7 RNA polymerase was observed in various mammalian cell lines after infection with the recombinant baculoviruses. After transfection of plasmids containing the luciferase gene under the control of the T7 promoter, high luciferase activity was detected in cells infected with the recombinant baculoviruses. We also constructed a plasmid containing an entire cDNA clone of type 1 poliovirus under the T7 promoter. Two days after transfection of the plasmid into the cells infected with the recombinant baculoviruses, a high titer of poliovirus was recovered. The use of the recombinant baculoviruses did not cause any cytopathic effects even at a high multiplicity of infection. The lack of replication ability and low toxicity are the advantageous features of the hybrid baculovirus-T7 polymerase system in comparison with the widely used vaccinia-T7 polymerase system for gene expression and recovery of infectious viruses from its cDNA. SN - 0042-6822 UR - https://www.unboundmedicine.com/medline/citation/9168881/A_hybrid_baculovirus_T7_RNA_polymerase_system_for_recovery_of_an_infectious_virus_from_cDNA_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0042-6822(97)98537-8 DB - PRIME DP - Unbound Medicine ER -