Packaging of AeDNV-GFP transducing virus by expression of densovirus structural proteins from a sindbis virus expression system.

Abstract:

:Genetic recombination resulting in the production of wild-type infectious virus is an obstacle in the current system for producing densovirus transducing particles. In order to eliminate this problem, a double subgenomic Sindbis virus (TE/3'2J/VP) was engineered that expresses the structural proteins (VPs) of Aedes densonucleosis virus (AeDNV) from the second subgenomic promoter. Expression of AeDNV VPs from TE/3'2J/VP was confirmed by Northern analysis of RNA from infected C6/36 (Aedes albopictus) cells and by indirect immunofluorescence in infected C6/36 cells and BHK-21 cells. TE/3'2J/VP was used to infect C6/36 cells transfected with p7NS1-GFP, a plasmid expressing the nonstructural genes of AeDNV and green fluorescent protein (GFP) as a reporter gene. This infection resulted in the production of AeDNV-GFP transducing virus, which is infectious to C6/36 cells and Aedes aegypti larvae, as determined by GFP expression. The TE/3'2J/VP packaging system produced titers of transducing virus comparable to those produced by the standard two-plasmid method. The possibility of recombination resulting in wild-type infectious virus in transducing densovirus stocks was eliminated by employing an RNA virus expression system to supply AeDNV structural proteins.

journal_name

Virology

journal_title

Virology

authors

Allen-Miura TM,Afanasiev BN,Olson KE,Beaty BJ,Carlson JO

doi

10.1006/viro.1999.9622

subject

Has Abstract

pub_date

1999-04-25 00:00:00

pages

54-61

issue

1

eissn

0042-6822

issn

1096-0341

pii

S0042-6822(99)99622-8

journal_volume

257

pub_type

杂志文章

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