- Page Contents:
- Applications
- Features
- What you provide
- What is included
- Advantages
- Schedule
- Notice
Construction of Adenovirus Vectors Containing One Expression Cassette in the E1 Region or the E3 Region
Applications:
- The most classic adenovirus vector (also known as first-generation)
- Transient gene expression in vitro, especially in cell lines that are refractory to other transfection methods
- Transient gene expression in vivo
Features:
- AdenoQuick™ or AdenoZap™
- Ad5 backbone
- E1 deletion 3.1 kb (bp 354-3503)
- Wild type or deleted E3 region (2.7 kb, corresponding to bp 28133-30818 in the Ad5 genome)
- Maximum size of the transgene is approximately 5 kb (WT E3)and 7.8 kb (ΔE3)
You provide:
- An expression cassette including a promoter, coding sequence and polyadenylation signal
- A restriction map, or preferably the sequence of your transgene
The service includes:
- Construction of a shuttle vector containing your expression cassette
- (AdenoQuick™ system only: construction of a cosmid containing the entire sequence of your recombinant virus)
- Virus recovery by transfecting the DNA into helper cells
- Virus clone isolation by plaque assay
- Confirmation of virus identity at the DNA level
You can customize your request: for example, you can decide to construct the shuttle vector yourself and send it to us for virus recovery. Another popular option is to allow O.D.260 Inc. to construct a cosmid which contains the entire sequence of your recombinant adenovirus and you will recover the virus yourself.
TopAdvantages of using our service:
- Multiple cloning systems available to suit your application (AdenoQuick™ or AdenoZap™)
- Short turn-around
- Cost-effective, especially if using the AdenoZap™ system
Estimated Timetable
| Minimum time to completion | |
| Service | Time |
|---|---|
| Shuttle vector construction | 4 days |
Cosmid construction | 4 days |
| Virus recovery (from transfection to virus plaque isolation) | 6 days |
| Confirmation of virus genome | 5 days |
| Total full service | 3 weeks |
An Important Note on Handling Adenovirus:
Adenovirus is an infectious respiratory virus and its use as a genetic vector requires the use of adequate containment equipment and practices. Biosafety Level 2 (BL2) is appropriate for most customers; however, for some, use of BL3 or higher level containment facility may be required. PLEASE NOTE, O.D.260 Inc. will not carry out projects that require BL3 (or above) containment.
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