In-Fusion™ assembly: seamless engineering of multidomain fusion proteins, modular vectors, and mutations
In-Fusion™ can join any two pieces of DNA that have a 15-bp overlap at their ends. The result is equivalent to a recombination event at the ends of the DNAs. The 15-bp overlap may be engineered by inclusion in primers used to PCR amplify a segment of DNA. Originally described for inserting one piece...
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| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2007-09-01
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| Series: | BioTechniques |
| Online Access: | https://www.future-science.com/doi/10.2144/000112536 |
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| _version_ | 1850152392791687168 |
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| author | Baogong Zhu Guifang Cai Emily O. Hall Gordon J. Freeman |
| author_facet | Baogong Zhu Guifang Cai Emily O. Hall Gordon J. Freeman |
| author_sort | Baogong Zhu |
| collection | DOAJ |
| description | In-Fusion™ can join any two pieces of DNA that have a 15-bp overlap at their ends. The result is equivalent to a recombination event at the ends of the DNAs. The 15-bp overlap may be engineered by inclusion in primers used to PCR amplify a segment of DNA. Originally described for inserting one piece of DNA into a restriction enzyme-digested plasmid, We have found In-Fusion can join four or more pieces of DNA in a single reaction. We used this insight to construct seamless fusion proteins, modular vectors with readily interchangeable segments, and novel mutagenesis strategies. Replacement In-Fusion can be used to delete any desired DNA segment in a plasmid and replace it with any desired new DNA segment without limitations on position or size. |
| format | Article |
| id | doaj-art-6575d7a712554d2ebe6243cda7f109bb |
| institution | OA Journals |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 2007-09-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj-art-6575d7a712554d2ebe6243cda7f109bb2025-08-20T02:25:59ZengTaylor & Francis GroupBioTechniques0736-62051940-98182007-09-0143335435910.2144/000112536In-Fusion™ assembly: seamless engineering of multidomain fusion proteins, modular vectors, and mutationsBaogong Zhu0Guifang Cai1Emily O. Hall2Gordon J. Freeman31Dana-Farber Cancer Institute Harvard Medical School, Boston, MA, USA1Dana-Farber Cancer Institute Harvard Medical School, Boston, MA, USA1Dana-Farber Cancer Institute Harvard Medical School, Boston, MA, USA1Dana-Farber Cancer Institute Harvard Medical School, Boston, MA, USAIn-Fusion™ can join any two pieces of DNA that have a 15-bp overlap at their ends. The result is equivalent to a recombination event at the ends of the DNAs. The 15-bp overlap may be engineered by inclusion in primers used to PCR amplify a segment of DNA. Originally described for inserting one piece of DNA into a restriction enzyme-digested plasmid, We have found In-Fusion can join four or more pieces of DNA in a single reaction. We used this insight to construct seamless fusion proteins, modular vectors with readily interchangeable segments, and novel mutagenesis strategies. Replacement In-Fusion can be used to delete any desired DNA segment in a plasmid and replace it with any desired new DNA segment without limitations on position or size.https://www.future-science.com/doi/10.2144/000112536 |
| spellingShingle | Baogong Zhu Guifang Cai Emily O. Hall Gordon J. Freeman In-Fusion™ assembly: seamless engineering of multidomain fusion proteins, modular vectors, and mutations BioTechniques |
| title | In-Fusion™ assembly: seamless engineering of multidomain fusion proteins, modular vectors, and mutations |
| title_full | In-Fusion™ assembly: seamless engineering of multidomain fusion proteins, modular vectors, and mutations |
| title_fullStr | In-Fusion™ assembly: seamless engineering of multidomain fusion proteins, modular vectors, and mutations |
| title_full_unstemmed | In-Fusion™ assembly: seamless engineering of multidomain fusion proteins, modular vectors, and mutations |
| title_short | In-Fusion™ assembly: seamless engineering of multidomain fusion proteins, modular vectors, and mutations |
| title_sort | in fusion™ assembly seamless engineering of multidomain fusion proteins modular vectors and mutations |
| url | https://www.future-science.com/doi/10.2144/000112536 |
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