Adenoviral System 3 (Tet-On 3G Inducible)

The Adeno-X Tet-On 3G systems generate very high fold induction, with up to 3,000-fold difference between induced and uninduced states

The Adeno-X Tet-On 3G systems generate very high fold-induction, with up to 3,000-fold difference between induced and uninduced states. Using equal amounts of high-titer supernatants, HeLa cells cultured at the indicated concentrations of Dox were infected with Adeno-X Tet-On 3G luciferase virus. Cultures were harvested and assayed for luciferase activity.

Expression increases with higher MOIs

Expression increases with higher MOIs. HeLa cells were infected with varying MOIs of pAdenoX Tet-On 3G adenovirus that expresses luciferase. After four hours, the media was replaced with fresh media +/- doxycycline (1 μg/ml). Cultures were harvested and assayed for luciferase activity. Maximal expression increases with increasing MOI, which also results in a slight increase in background expression.

The Adeno-X Tet-On 3G systems are highly inducible

The Adeno-X Tet-On 3G systems are highly inducible. Using equal amounts of high-titer supernatants, HeLa cells cultured at the indicated concentrations of Dox were infected with Adeno-X Tet-On 3G lacZ virus. Cultures were harvested and assayed for beta-galactosidase activity using the Luminescent Beta-galactosidase Reporter System 3 (Cat. # 631713).

The Adeno-X Adenoviral System 3 (Tet-On 3G Inducible) allows inducible gene expression only in the presence of doxycycline

The Adeno-X Adenoviral System 3 (Tet-On 3G Inducible) allows inducible gene expression only in the presence of doxycycline. The system includes In-Fusion HD Cloning Kit for cloning your gene of interest (GOI) directly into the easy-to-use, all-in-one pAdenoX-Tet3G expression vector.

Comparison of Adeno-X adenoviral system 3 to the leading “Easy” competitor

Comparison of Adeno-X adenoviral system 3 to the leading “Easy” competitor.

Constructing recombinant adenovirus with In-Fusion Cloning technology

Constructing recombinant adenovirus with In-Fusion Cloning technology. DNA sequences can be rapidly transferred as PCR products to any pAdenoX vector using the In-Fusion cloning method. In this example, your gene of interest is amplified with 15 bp extensions that are homologous to the ends of the linearized adenoviral vector. The PCR product is then purified and mixed with the linearized adenoviral vector of choice in the In-Fusion reaction. Following the reaction, a portion of the mixture is transformed into E. coli (Stellar Competent Cells) and screened. Once a PCR-positive clone is identified, the recombinant pAdenoX vector is amplified, purified, and subsequently linearized with the restriction enzyme PacI, then transfected into Adeno-X 293 cells for viral rescue and amplification. Adeno-X GoStix can be used to determine the status of adenovirus rescue.