Secretory fluorescent protein, a secretion green fluorescent fusion protein with alkaline phosphatase activity as a sensitive and traceable reporter in baculovirus expression system
| Autor: | Chao-Yi Teng, Tzong-Yuan Wu |
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| Rok vydání: | 2007 |
| Předmět: |
Reporter gene
Brefeldin A Recombinant Fusion Proteins Green Fluorescent Proteins Gene Expression Bioengineering General Medicine Biology Alkaline Phosphatase Applied Microbiology and Biotechnology Molecular biology Fusion protein Cell Line Green fluorescent protein chemistry.chemical_compound Secretory protein chemistry Genes Reporter Animals Alkaline phosphatase Secretion Baculoviridae Secretory pathway Biotechnology |
| Zdroj: | Biotechnology Letters. 29:1019-1024 |
| ISSN: | 1573-6776 0141-5492 |
| DOI: | 10.1007/s10529-007-9349-y |
| Popis: | The advantages of using traceable fluorescent protein (enhanced green fluorescent protein; EGFP) and a secretory alkaline phosphatase (SEAP) have been used to generate a reporter gene: the secretory fluorescent protein (SEFP). Sf21 cells, infected with the recombinant baculovirus containing the SEFP gene, revealed both traceable fluorescence and easily detectable alkaline phosphatase activity in the culture medium. The distribution of SEFP within the cells revealed that it was excluded from the nucleus, implying that the accumulation of SEFP in a secretory pathway, similar to that of the secretion signal-tagged FPs. Furthermore, the time- and dose-dependent release from the blockage of brefeldin A (BFA) confirmed that the secretion of SEFP was mediated by the secretion pathway and excluded leakage from viral infection. This SEFP reporter gene with traceable fluorescence and alkaline phosphatase activity may become a useful tool for studies on secretory protein production. |
| Databáze: | OpenAIRE |
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