Simultaneous Insertion of Two Expression Cassettes into Adenovirus Vectors
We have designed AdenoQuick, a fast and versatile method to construct first-generation adenoviral vectors that contain one or two transgenes in the E1 and/or the E3 region. The method is based on the reconstitution of the entire genome of the desired recombinant virus in E. coli and the subsequent t...
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| Main Author: | |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2001-03-01
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| Series: | BioTechniques |
| Online Access: | https://www.future-science.com/doi/10.2144/01303rr06 |
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| Summary: | We have designed AdenoQuick, a fast and versatile method to construct first-generation adenoviral vectors that contain one or two transgenes in the E1 and/or the E3 region. The method is based on the reconstitution of the entire genome of the desired recombinant virus in E. coli and the subsequent transfection of the DNA in a helper cell line. Since the construction of large adenoviral plasmids is generally difficult and therefore rebuffing for inexperienced researchers, we have optimized the cloning strategy by using bacterial positive-selection markers and a set of specific restriction enzymes that allow for directional cloning. The system is 99% efficient and allows one to insert simultaneously two expression cassettes into the E1 and E3 regions of the adenovirus genome. |
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| ISSN: | 0736-6205 1940-9818 |