Optimization of bovine coronavirus hemagglutinin-estrase glycoprotein expression in E3 deleted bovine adenovirus-3
Identifieur interne : 000058 ( France/Analysis ); précédent : 000057; suivant : 000059Optimization of bovine coronavirus hemagglutinin-estrase glycoprotein expression in E3 deleted bovine adenovirus-3
Auteurs : P. Seshidhar Reddy [Canada] ; Neeraja Idamakanti [Canada] ; Lexander N. Zakhartchouk [Canada] ; Lorne A. Babiuk [Canada] ; Majid Mehtali [France] ; Suresh K. Tikoo [Canada]Source :
- Virus Research [ 0168-1702 ] ; 2000.
English descriptors
- KwdEn :
- Teeft :
- Adenovirus, Babiuk, Bamhi, Bamhi fragment, Bovine, Bovine adenovirus type, Bovine coronavirus, Cdna, Cdna sequence, Cell lysates, Chimeric, Chimeric intron, Coli, Coronavirus, Cotton rats, Exogenous, Exogenous transcription control elements, Expression system, Foreign genes, Gene expression, Genome, Glycoprotein, Idamakanti, Immunoprecipitation analysis, Intron, Kpni, Kpni spei fragment, Late signal, Mdbk, Mdbk cells, Monoclonal antibodies, Mrna, Nytran membranes, Plasmid, Plasmid psvpia, Post infection, Postinfection, Promoter, Recombinant, Recombinant genomes, Recombinant viruses, Recombination, Reddy, Right side, Srfi, Srfi linearized plasmid, Tikoo, Transcription, Transfer vector, Virol, Virus research, Zakhartchouk.
Abstract
Abstract: Adenoviral vectors expressing foreign genes have many desirable properties in applications such as vaccination. Recently, we have generated replication-competent (E3 deleted) bovine adenovirus-3 (BAV-3) recombinants expressing significant amounts of glycoprotein D (gD) of bovine herpesvirus-1 (a DNA virus). However, attempts to express the RNA virus genes using the same strategy were not successful. In an effort to optimize the expression, we have constructed several BAV-3 recombinants carrying the hemagglutinin esterase (HE) gene of bovine coronavirus (BCV) in the E3 region with or without exogenous transcription control elements. The expression studies suggest that the introduction of a 137 bp chimeric intron upstream of the HE cDNA is able to increase the level of HE gene expression. The introduction of a SV40 early promoter or human cytomegalovirus (HCMV) immediate early (IE) promoter into the expression cassette changed the kinetics of the HE expression. However, the recombinant BAV-3 containing HE under the HCMV IE promoter replicated less efficiently than the wild-type BAV-3. These studies should prove useful in expression of other RNA viral genes in the E3 region of BAV-3 expression system.
Url:
DOI: 10.1016/S0168-1702(00)00209-4
Affiliations:
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<front><div type="abstract" xml:lang="en">Abstract: Adenoviral vectors expressing foreign genes have many desirable properties in applications such as vaccination. Recently, we have generated replication-competent (E3 deleted) bovine adenovirus-3 (BAV-3) recombinants expressing significant amounts of glycoprotein D (gD) of bovine herpesvirus-1 (a DNA virus). However, attempts to express the RNA virus genes using the same strategy were not successful. In an effort to optimize the expression, we have constructed several BAV-3 recombinants carrying the hemagglutinin esterase (HE) gene of bovine coronavirus (BCV) in the E3 region with or without exogenous transcription control elements. The expression studies suggest that the introduction of a 137 bp chimeric intron upstream of the HE cDNA is able to increase the level of HE gene expression. The introduction of a SV40 early promoter or human cytomegalovirus (HCMV) immediate early (IE) promoter into the expression cassette changed the kinetics of the HE expression. However, the recombinant BAV-3 containing HE under the HCMV IE promoter replicated less efficiently than the wild-type BAV-3. These studies should prove useful in expression of other RNA viral genes in the E3 region of BAV-3 expression system.</div>
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