Construction of a full-length cDNA of tobacco mosaic virus strain V-69 genome

Construction of a full-length cDNA of tobacco mosaic virus strain V-69 genome The genome of the tobacco mosaic virus (TMV) strain V-69 was sequenced by us earlier. The comparison of its nucleotide sequence with those of well-known tobamovirus strains (using published data) demonstrated considerable homology between strain V-69 and tomato strain L of TMV. In this study, full-length cDNA copy of TMV strain V-69 genomic RNA has been cloned in vector pBS(+) under the control of the T7 promoter and with an SmaI site at the 3’-terminus. The in vitro transcription of this construction with the use of phage T7 RNA polymerase in the presence of a cap analog (m7 GpppG) has yielded an infectious RNA. This RNA has induced the same symptoms as those caused by TMV strain V-69 in infected indicator plants. Three point mutations inactivating one of two HindIII sites have been introduced into the cDNA. The absence of the HindIII site at position 1449 of the viral nucleotide sequence and the corresponding mutations serve as markers of the construction obtained. The full-length cDNA copy of the TMV strain V-69 RNA can be used both as a research tool and for constructing a vector for foreign gene expression. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Russian Journal of Genetics Springer Journals

Construction of a full-length cDNA of tobacco mosaic virus strain V-69 genome

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Publisher
Nauka/Interperiodica
Copyright
Copyright © 2004 by MAIK “Nauka/Interperiodica”
Subject
Biomedicine; Human Genetics; Microbial Genetics and Genomics; Animal Genetics and Genomics
ISSN
1022-7954
eISSN
1608-3369
D.O.I.
10.1007/s11177-005-0058-0
Publisher site
See Article on Publisher Site

Abstract

The genome of the tobacco mosaic virus (TMV) strain V-69 was sequenced by us earlier. The comparison of its nucleotide sequence with those of well-known tobamovirus strains (using published data) demonstrated considerable homology between strain V-69 and tomato strain L of TMV. In this study, full-length cDNA copy of TMV strain V-69 genomic RNA has been cloned in vector pBS(+) under the control of the T7 promoter and with an SmaI site at the 3’-terminus. The in vitro transcription of this construction with the use of phage T7 RNA polymerase in the presence of a cap analog (m7 GpppG) has yielded an infectious RNA. This RNA has induced the same symptoms as those caused by TMV strain V-69 in infected indicator plants. Three point mutations inactivating one of two HindIII sites have been introduced into the cDNA. The absence of the HindIII site at position 1449 of the viral nucleotide sequence and the corresponding mutations serve as markers of the construction obtained. The full-length cDNA copy of the TMV strain V-69 RNA can be used both as a research tool and for constructing a vector for foreign gene expression.

Journal

Russian Journal of GeneticsSpringer Journals

Published: Feb 16, 2005

References

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