Adenovirus Construction Services

Techniques Introduction:

Recombinant adenovirus is one of the most efficient and reliable recombinant viral expression systems available for transient expression of shRNA or target proteins in mammalian cells at instantaneously and high levels.

There is no question that the adenovirus is the most effective means of delivering genes in vivo and in vitro. Construction of adenovirus is time-consuming and labor intensive. 

Advantages：
    – Host cell range: Adenovirus can infect a range of mammalian cells, in addition to infect almost all human cell types, but also infected rats, mice, rabbits, pigs, sheep, monkeys, chickens and other species.
    – Because adenovirus infection does not depend on the cell cycle, it can infect both proliferating cells and no proliferative cells.
    – Transient expression, do not need to be integrated into the genome.
    – The virus titer is high and can be used for animal level experiments.

The system is based on Ad5 serotype E1 / E3 deficiency adenovirus DNA as the skeleton, improve and optimize in the cloning, packaging, amplification and other conditions to achieve a higher positive rate. It is more reliable and with advantages like, stable packaging, poisons fast and high titer effect. The packaged adenovirus has been successfully applied to apoptosis detection, chromatin ChIP, RT-PCR, Co-IP and other scientific research experiments.

Experimental Steps and Cycles:
    – The target gene was inserted into the adenovirus re-shuttle plasmid, the experimental period was 7 working days, and the experimental route and the experimental cycle were specified when the specific parts of the carrier were required to be specially modified.
    – It will take 7 working days when the target gene was transferred to the adenovirus backbone.
    – The correct recombinant adenovirus plasmid was identified in 293A cells for packaging for approximately 20 working days.
    – Adenovirus amplification and CsCl purification, the experimental period of 7 working days.
    – The quality of the packaged virus is packaged, including PCR and WB of the final product virus gene to confirm the presence of the target gene, with an experimental cycle of 5 working days.

Pledge:
    – Provided the amount of virus for 1ml titer> 1010VP / ml, and a greater amount ones, please contact the specific.
    – An electrophoresis map, sequencing report, final product PCR or WB identification of shuttle plasmid, recombinant adenovirus plasmid and related enzyme digestion were provided.
    – In order to ensure the quality of service, the contract cycle is based on the experimental cycle plus 10 working days.

Applications:  

Fig1: Infection efficiency of Ad-Fos in CGNs. DIV5 CGNs was infected with Ad-Fos at MOI of 100. Thirty six hours after infection phase-contrast light and fluorescent micrographs of the infected cells were visualized. The cells infected with Ad-Fos were fixed and stained with a Flag-specific M2 antibody or without a primary antibody (-1°). Infection efficiency of adenovirus is about 80%.