Adenoviruses are medium-sized (80–100nm), non-enveloped viruses. They have an icosahedral nucleocapsid containing a linear, double-stranded DNA genome of approximately 36kb (Nermut, 1984). The viral genome is grouped into different transcriptional units, designated early (E1, E2, E3, E4), intermediate, and late. The E1 gene is essential for activation of other viral genes and for viral replication. Deletion of the E1 gene results in viruses that are replication incompetent in normal cells. However, replication-competent viral particles can be produced from E1-deleted viral vectors by providing the E1 gene in trans. The E3 gene is nonessential for either viral replication or infection (Flint, 1999). Adenovirus type 5 is one of the most extensively studied and characterized adenoviruses and is the type used most frequently in generating recombinant adenoviruses for gene therapy. These vectors generally contain deletions of the E1 and E3 genes, which allows for insertion and packaging of up to 7.5kb of foreign DNA, for gene delivery. This recombinant human adenovirus type 5 expresses beta-galactosidase protein under the control of a CMV promoter. beta-galactosidase is a glycoside hydrolase enzyme that catalyzes the hydrolysis of beta-galactosides into monosaccharides through the breaking of a glycosidic bond. An active enzyme may be detected using artificial chromogenic substrate 5-bromo-4-chloro-3-indolyl-beta-d-galactopyranoside, X-gal. beta-galactosidase will cleave the glycosidic bond in X-gal and form galactose and 5-bromo-4-chloro-3-hydroxyindole which dimerizes and oxidizes to 5,5′-dibromo-4,4′-dichloro-indigo, producing an intense blue precipitate. Therefore, this vector can be used to determine virus localisation, transduction efficiency and to optimize viral infection conditions in specific cell types.
Description:Ad5 with an E1 and E3 deletion. Insertion of CMV-driven nuclear-targeted β-galatosidase gene in E1 region. Concentrated and purified virus particles from a double CsCl gradient purification with DNase treatment and dialysis.
Cells Grown In:293AD cells
Storage and Stability:Aliquot to avoid repeated freezing and thawing and store at -70°C. Aliquots are stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
仕様
Size：100ul
Source Antigen：Native
Grade：Highly Purified
Purity：≥99%
Form：Supplied as a liquid in 50mM HEPES, pH 7.8, 7.5% sucrose.
