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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Szymczak-Workman, Andrea L. Vignali, Kate M. Vignali, Dario A. A. |
| Abstract | The need for reliable, multicistronic vectors for multigene delivery is at the forefront of biomedical technology. It is now possible to express multiple proteins from a single open reading frame (ORF) using 2A peptide-linked multicistronic vectors. These small sequences, when cloned between genes, allow for efficient, stoichiometric production of discrete protein products within a single vector through a novel 'cleavage' event within the 2A peptide sequence. The easiest and most effective way to assess 2A cleavage is to perform transient transfection of 293T cells (human embryonic kidney cells) followed by western blot analysis, as described in this protocol. 293T cells are easy to grow and can be efficiently transfected with a variety of vectors. Cleavage can be assessed by detection with antibodies against the target proteins or anti-2A serum. |
| File Format | HTM / HTML |
| ISSN | 19403402 |
| Issue Number | 2 |
| Volume Number | 2012 |
| e-ISSN | 15596095 |
| Journal | Cold Spring Harbor Protocols |
| Language | English |
| Publisher | Cold Spring Harbor Laboratory Press |
| Publisher Date | 2012-02-01 |
| Publisher Place | United States |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Discipline Clinical Laboratory Techniques Genetic Engineering Methods Peptides Genetics Metabolism Proteolysis Blotting, Western Cell Line Gene Expression Genes Genetic Vectors Humans Open Reading Frames Protein Biosynthesis Protein Processing, Post-translational Ribosomes Transfection Journal Article |
| Content Type | Text |
| Resource Type | Article |
| Subject | Biochemistry, Genetics and Molecular Biology |
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