2019
DOI: 10.1073/pnas.1910787117
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Rescue of tomato spotted wilt virus entirely from complementary DNA clones

Abstract: Negative-stranded/ambisense RNA viruses (NSVs) include not only dangerous pathogens of medical importance but also serious plant pathogens of agronomic importance. Tomato spotted wilt virus (TSWV) is one of the most important plant NSVs, infecting more than 1,000 plant species, and poses major threats to global food security. The segmented negative-stranded/ambisense RNA genomes of TSWV, however, have been a major obstacle to molecular genetic manipulation. In this study, we report the complete recovery of inf… Show more

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Cited by 66 publications
(83 citation statements)
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“…Full‐length ALSV RNA1 (GenBank accession no. AB030940) and RNA2 (AB030941) sequences were synthesized by the GenScript (Nanjing, China) and cloned individually into the pCB301‐2 × 35S‐RZ‐NOS vector (Feng et al , 2020) to produce pALSV1 and pALSV2, respectively. A partial sequence (300 bp) of AtPME or AtGBL was RT‐PCR amplified from a total RNA sample using specific primers and cloned into the pALSV2 vector to produce pALSV2‐AtPME or pALSV2‐AtGBL.…”
Section: Methodsmentioning
confidence: 99%
“…Full‐length ALSV RNA1 (GenBank accession no. AB030940) and RNA2 (AB030941) sequences were synthesized by the GenScript (Nanjing, China) and cloned individually into the pCB301‐2 × 35S‐RZ‐NOS vector (Feng et al , 2020) to produce pALSV1 and pALSV2, respectively. A partial sequence (300 bp) of AtPME or AtGBL was RT‐PCR amplified from a total RNA sample using specific primers and cloned into the pALSV2 vector to produce pALSV2‐AtPME or pALSV2‐AtGBL.…”
Section: Methodsmentioning
confidence: 99%
“…These observations raise the intriguing question of whether these proteins can overcome the antisense problem to facilitate rescue of mammalian nonsegmented NSVs from genomic cDNA copies. For segmented NSVs of plants and some animal counterparts, the current reverse genetics system schemes involve transcription of viral agRNAs that serve as templates for both genome replication and translation of NC proteins [ 20 , 21 , 24 , 29 , 30 ]. In these cases, adaptation of the gRNA approach would require additional plasmids to provide the NC proteins, and these in trans expressed proteins may be less efficient in recruiting the RNA templates for NC assembly [ 52 ].…”
Section: Discussionmentioning
confidence: 99%
“…Similar strategies were applicable to a barley yellow striate mosaic cytorhabdovirus, although infections of its native monocot hosts require insect vector-mediated transfer of the recombinant virus recovered from N. benthamiana leaves [ 28 ]. Recently, refined positive-strand agRNA strategies were also adapted to rescue a tri-segmented tospovirus [ 29 ] and a seven-segmented emaravirus [ 30 ]. In these two cases, expression of viral core proteins (N and L) from in trans provided supporting plasmids is unnecessary since the agRNA transcripts permit direct ribosomal translation to produce both core proteins in cis [ 29 , 30 ].…”
Section: Introductionmentioning
confidence: 99%
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“…However, there is still no evidence that CCA1 influences TSWV infectivity. The first report of an infectious TSWV clone ( Feng et al, 2020 ) may help to elucidate whether CCA1 can interact with the native TSWV M RNA in planta .…”
Section: The Core Circadian Clock Component Cca1 Suppresses Photomorpmentioning
confidence: 99%