A time- and cost-efficient system for high-level protein production in mammalian cells.
Most proteins for structural biology studies are produced by high-level expression in Escherichia coli. However, prokaryotic based expression systems fail to generate correctly folded functional forms of many proteins and hence a variety of eukaryotic based expression systems have been developed. Of...
| Hlavní autoři: | , , |
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| Médium: | Journal article |
| Jazyk: | English |
| Vydáno: |
2006
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| _version_ | 1826283964905553920 |
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| author | Aricescu, A Lu, W Jones, E |
| author_facet | Aricescu, A Lu, W Jones, E |
| author_sort | Aricescu, A |
| collection | OXFORD |
| description | Most proteins for structural biology studies are produced by high-level expression in Escherichia coli. However, prokaryotic based expression systems fail to generate correctly folded functional forms of many proteins and hence a variety of eukaryotic based expression systems have been developed. Of these, yeast and baculovirus-infected insect cells currently represent the expression systems of choice for structural biologists. Here, protocols for a simple, fast and affordable method for transient protein expression in mammalian cells are reported. The results demonstrate that it combines several features necessary for the production of suitable samples for structural biology, in particular protein crystallography, namely high protein yield, straightforward purification, selenomethionine incorporation and control of N-linked glycosylation. The system is suitable for use in conventional laboratories or can be implemented in a medium- or high-throughput pipeline. |
| first_indexed | 2024-03-07T01:06:44Z |
| format | Journal article |
| id | oxford-uuid:8b9b13e0-0800-43e7-b935-84d5410086a9 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T01:06:44Z |
| publishDate | 2006 |
| record_format | dspace |
| spelling | oxford-uuid:8b9b13e0-0800-43e7-b935-84d5410086a92022-03-26T22:39:06ZA time- and cost-efficient system for high-level protein production in mammalian cells.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:8b9b13e0-0800-43e7-b935-84d5410086a9EnglishSymplectic Elements at Oxford2006Aricescu, ALu, WJones, EMost proteins for structural biology studies are produced by high-level expression in Escherichia coli. However, prokaryotic based expression systems fail to generate correctly folded functional forms of many proteins and hence a variety of eukaryotic based expression systems have been developed. Of these, yeast and baculovirus-infected insect cells currently represent the expression systems of choice for structural biologists. Here, protocols for a simple, fast and affordable method for transient protein expression in mammalian cells are reported. The results demonstrate that it combines several features necessary for the production of suitable samples for structural biology, in particular protein crystallography, namely high protein yield, straightforward purification, selenomethionine incorporation and control of N-linked glycosylation. The system is suitable for use in conventional laboratories or can be implemented in a medium- or high-throughput pipeline. |
| spellingShingle | Aricescu, A Lu, W Jones, E A time- and cost-efficient system for high-level protein production in mammalian cells. |
| title | A time- and cost-efficient system for high-level protein production in mammalian cells. |
| title_full | A time- and cost-efficient system for high-level protein production in mammalian cells. |
| title_fullStr | A time- and cost-efficient system for high-level protein production in mammalian cells. |
| title_full_unstemmed | A time- and cost-efficient system for high-level protein production in mammalian cells. |
| title_short | A time- and cost-efficient system for high-level protein production in mammalian cells. |
| title_sort | time and cost efficient system for high level protein production in mammalian cells |
| work_keys_str_mv | AT aricescua atimeandcostefficientsystemforhighlevelproteinproductioninmammaliancells AT luw atimeandcostefficientsystemforhighlevelproteinproductioninmammaliancells AT jonese atimeandcostefficientsystemforhighlevelproteinproductioninmammaliancells AT aricescua timeandcostefficientsystemforhighlevelproteinproductioninmammaliancells AT luw timeandcostefficientsystemforhighlevelproteinproductioninmammaliancells AT jonese timeandcostefficientsystemforhighlevelproteinproductioninmammaliancells |