Detection of expressed chloramphenicol acetyltransferase in the saliva of Culex pipiens mosquitoes.
Insect Biochem Mol Biol
; 27(5): 423-9, 1997 May.
Article
em En
| MEDLINE
| ID: mdl-9219368
ABSTRACT
Mosquito salivary glands play an important role in the transmission of arthropod-borne pathogens. The ability to express genes in mosquitoes would be a powerful approach to characterize salivary gland genes, and to reveal important vector determinants of pathogen transmission. Here we report the use of a double subgenomic Sindbis (dsSIN) virus, designated TE/3'2J/CAT, and a packaged Sindbis replicon virus, designated rep5/CAT/26S, to express chloramphenicol acetyltransferase (CAT) protein in the salivary glands and saliva of transduced female Culex pipiens pipiens. Indirect immunofluorescence analysis revealed that salivary glands of these mosquitoes infected with either TE/3'2J/CAT or rep5/CAT/26S virus (4 or 6 days post-infection (p.i.)) were positive for both SIN E1 antigen and CAT protein. Saliva collected from mosquitoes transduced with TE/3'2J/CAT virus contained a unique 25 kDa protein that corresponded to the size of CAT protein. Additionally, CAT activity assays revealed that saliva collected from mosquitoes transduced with either TE/3'2J/CAT or rep5/CAT/26S virus could contain greater than 5.0 x 10(-5) units of CAT enzyme (3.0 x 10(6) CAT trimers).
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Sindbis virus
/
Glicoproteínas de Membrana
/
Cloranfenicol O-Acetiltransferase
/
Proteínas do Envelope Viral
/
Culex
/
Vetores Genéticos
Tipo de estudo:
Diagnostic_studies
Limite:
Animals
Idioma:
En
Revista:
Insect Biochem Mol Biol
Assunto da revista:
BIOLOGIA MOLECULAR
/
BIOQUIMICA
Ano de publicação:
1997
Tipo de documento:
Article
País de afiliação:
Estados Unidos