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| Format: | Artículo científico |
| Language: | en |
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Academia Brasileira de Ciências
2002
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| Online Access: | https://www.redalyc.org/articulo.oa?id=32774405 |
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| _version_ | 1866812394185949184 |
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| author | Marcelo Surpili |
| author_facet | Marcelo Surpili |
| contents | A yeast-based model system for cloning secreted and membrane proteins Marcelo Surpili Bernd Müller Röber Lothar Willmitzer Multidisciplinaria (Ciencias Naturales y Exactas) signal sequences potassium channel library screening yeast acid phosphatase The targeting of proteins to cell organelles and membranes, or of proteins destined to secretion, is coordinatedby signal sequences located at the 5´-end of their respective genes. A signal sequence trap system wasenvisaged in which a truncated version of the yeast acid phosphatase pho5 gene lacking the start codonand signal sequence could serve as a reporter gene. A fraction enriched in 5´-end fragments obtained byPCR from a potato guard-cell cDNA library was cloned in frame to the acid phosphatase gene and the acidphosphatase activity was assayed directly in yeast colonies grown on selective medium. Putative signalsequences targeting the acid phosphatase to the membrane or to the outside of the cell were used to screen thecDNA bank in order to recover the original full-size sequence which gave rise to the signal sequence. Twounknown sequences displaying marked tissue-specific expression were retrieved, one of them (YE139) witha higher expression level in green buds and stem cells, and the other one (YE290) with a higher expressionlevel in androceum, gyneceum, and roots. The limitations of the system are further analyzed using othersequences as control. 2002 artículo científico 0001-3765 https://www.redalyc.org/articulo.oa?id=32774405 en http://www.redalyc.org/revista.oa?id=327 Anais da Academia Brasileira de Ciências application/pdf Academia Brasileira de Ciências Anais da Academia Brasileira de Ciências (Brasil) Num.4 Vol.74 |
| format | Artículo científico |
| id | redalyc_32774405 |
| language | en |
| publishDate | 2002 |
| publisher | Academia Brasileira de Ciências |
| spellingShingle | A yeast-based model system for cloning secreted and membrane proteins Marcelo Surpili Multidisciplinaria (Ciencias Naturales y Exactas) signal sequences potassium channel library screening yeast acid phosphatase A yeast-based model system for cloning secreted and membrane proteins Marcelo Surpili Bernd Müller Röber Lothar Willmitzer Multidisciplinaria (Ciencias Naturales y Exactas) signal sequences potassium channel library screening yeast acid phosphatase The targeting of proteins to cell organelles and membranes, or of proteins destined to secretion, is coordinatedby signal sequences located at the 5´-end of their respective genes. A signal sequence trap system wasenvisaged in which a truncated version of the yeast acid phosphatase pho5 gene lacking the start codonand signal sequence could serve as a reporter gene. A fraction enriched in 5´-end fragments obtained byPCR from a potato guard-cell cDNA library was cloned in frame to the acid phosphatase gene and the acidphosphatase activity was assayed directly in yeast colonies grown on selective medium. Putative signalsequences targeting the acid phosphatase to the membrane or to the outside of the cell were used to screen thecDNA bank in order to recover the original full-size sequence which gave rise to the signal sequence. Twounknown sequences displaying marked tissue-specific expression were retrieved, one of them (YE139) witha higher expression level in green buds and stem cells, and the other one (YE290) with a higher expressionlevel in androceum, gyneceum, and roots. The limitations of the system are further analyzed using othersequences as control. 2002 artículo científico 0001-3765 https://www.redalyc.org/articulo.oa?id=32774405 en http://www.redalyc.org/revista.oa?id=327 Anais da Academia Brasileira de Ciências application/pdf Academia Brasileira de Ciências Anais da Academia Brasileira de Ciências (Brasil) Num.4 Vol.74 |
| title | A yeast-based model system for cloning secreted and membrane proteins |
| topic | Multidisciplinaria (Ciencias Naturales y Exactas) signal sequences potassium channel library screening yeast acid phosphatase |
| url | https://www.redalyc.org/articulo.oa?id=32774405 |