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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| 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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| _version_ | 1850152941429719040 |
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| author | Xavier Danthinne |
| author_facet | Xavier Danthinne |
| author_sort | Xavier Danthinne |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-a69e4379aa2e4250954d27429fcd494b |
| institution | OA Journals |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 2001-03-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj-art-a69e4379aa2e4250954d27429fcd494b2025-08-20T02:25:51ZengTaylor & Francis GroupBioTechniques0736-62051940-98182001-03-0130361261910.2144/01303rr06Simultaneous Insertion of Two Expression Cassettes into Adenovirus VectorsXavier Danthinne01O.D. 260 Inc., Mountain States, Medical Research Institute, and Department of Veterans Affairs, Medical Center, Boise, ID, USAWe 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.https://www.future-science.com/doi/10.2144/01303rr06 |
| spellingShingle | Xavier Danthinne Simultaneous Insertion of Two Expression Cassettes into Adenovirus Vectors BioTechniques |
| title | Simultaneous Insertion of Two Expression Cassettes into Adenovirus Vectors |
| title_full | Simultaneous Insertion of Two Expression Cassettes into Adenovirus Vectors |
| title_fullStr | Simultaneous Insertion of Two Expression Cassettes into Adenovirus Vectors |
| title_full_unstemmed | Simultaneous Insertion of Two Expression Cassettes into Adenovirus Vectors |
| title_short | Simultaneous Insertion of Two Expression Cassettes into Adenovirus Vectors |
| title_sort | simultaneous insertion of two expression cassettes into adenovirus vectors |
| url | https://www.future-science.com/doi/10.2144/01303rr06 |
| work_keys_str_mv | AT xavierdanthinne simultaneousinsertionoftwoexpressioncassettesintoadenovirusvectors |