About: A Reverse Genetics System for Cypovirus Based on a Bacmid Expressing T7 RNA Polymerase

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type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
title	A Reverse Genetics System for Cypovirus Based on a Bacmid Expressing T7 RNA Polymerase
Creator	Yang, Jian Hu, Jia Wang, Jia Lei, Chengfeng Qin, Fujun »more»
source	Medline; PMC
abstract	Dendrolimus punctatus cypovirus (DpCPV), belonging to the genus Cypovirus within the family Reoviridae, is considered the most destructive pest of pine forests worldwide. DpCPV has a genome consisting of 10 linear double-stranded RNA segments. To establish a reverse genetics system, we cloned cDNAs encoding the 10 genomic segments of DpCPV into three reverse genetics vectors in which each segment was transcribed under the control of a T7 RNA polymerase promoter and terminator tagged with a hepatitis delta virus ribozyme sequence. We also constructed a vp80-knockout Autographa californica multiple nucleopolyhedrovirus bacmid to express a T7 RNA polymerase codon-optimized for Sf9 cells. Following transfection of Sf9 cells with the three vectors and the bacmid, occlusion bodies (OBs) with the typical morphology of cypovirus polyhedra were observed by optical microscopy. The rescue system was verified by incorporation of a HindIII restriction enzyme site null mutant of the 9th genomic segment. Furthermore, when we co-transfected Sf9 cells with the reverse genetics vectors, the bacmid, and an additional vector bearing an egfp gene flanked with the 5′ and 3′ untranslated regions of the 10th genomic segment, aggregated green fluorescence co-localizing with the OBs was observed. The rescued OBs were able to infect Spodopetra exigua larvae, although their infectivity was significantly lower than that of wild-type DpCPV. This reverse genetics system for DpCPV could be used to explore viral replication and pathogenesis and to facilitate the development of novel bio-insecticides and expression systems for exogenous proteins.
has issue date	2019-04-01(xsd:dateTime)
bibo:doi	10.3390/v11040314
bibo:pmid	30939777
has license	cc-by
sha1sum (hex)	b37857f5ea12c3dadcd0929c3c38720fc8c7d028
schema:url	https://doi.org/10.3390/v11040314
resource representing a document's title	A Reverse Genetics System for Cypovirus Based on a Bacmid Expressing T7 RNA Polymerase
has PubMed Central identifier	PMC6521135
has PubMed identifier	30939777
schema:publication	Viruses
resource representing a document's body	covid:b37857f5ea12c3dadcd0929c3c38720fc8c7d028#body_text
is schema:about of	named entity 'LARVAE' named entity 'FAMILY REOVIRIDAE' named entity 'GREEN ' named entity 'GENOMIC' named entity 'ESTABLISH' »more»

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