Medium-throughput production of recombinant human proteins: ligation-independent cloning.
Structural genomics groups have identified the need to generate multiple truncated versions of each target to improve their success in producing a well-expressed, soluble, and stable protein and one that crystallizes and diffracts to a sufficient resolution for structural determination. At the SGC,...
| Main Authors: | , , , |
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| Format: | Journal article |
| Language: | English |
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2014
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| _version_ | 1826274153000337408 |
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| author | Strain-Damerell, C Mahajan, P Gileadi, O Burgess-Brown, N |
| author_facet | Strain-Damerell, C Mahajan, P Gileadi, O Burgess-Brown, N |
| author_sort | Strain-Damerell, C |
| collection | OXFORD |
| description | Structural genomics groups have identified the need to generate multiple truncated versions of each target to improve their success in producing a well-expressed, soluble, and stable protein and one that crystallizes and diffracts to a sufficient resolution for structural determination. At the SGC, we opted for the Ligation-Independent Cloning (LIC) method which provides the medium throughput we desire to produce and screen many proteins in a parallel process. Here, we describe our LIC protocol for generating constructs in a 96-well format and provide a choice of vectors suitable for expressing proteins in both E. coli and the baculovirus expression vector system (BEVS). |
| first_indexed | 2024-03-06T22:39:07Z |
| format | Journal article |
| id | oxford-uuid:5aeb3b52-e76c-4e82-b872-d2ffdfe674bf |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T22:39:07Z |
| publishDate | 2014 |
| record_format | dspace |
| spelling | oxford-uuid:5aeb3b52-e76c-4e82-b872-d2ffdfe674bf2022-03-26T17:18:51ZMedium-throughput production of recombinant human proteins: ligation-independent cloning.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:5aeb3b52-e76c-4e82-b872-d2ffdfe674bfEnglishSymplectic Elements at Oxford2014Strain-Damerell, CMahajan, PGileadi, OBurgess-Brown, NStructural genomics groups have identified the need to generate multiple truncated versions of each target to improve their success in producing a well-expressed, soluble, and stable protein and one that crystallizes and diffracts to a sufficient resolution for structural determination. At the SGC, we opted for the Ligation-Independent Cloning (LIC) method which provides the medium throughput we desire to produce and screen many proteins in a parallel process. Here, we describe our LIC protocol for generating constructs in a 96-well format and provide a choice of vectors suitable for expressing proteins in both E. coli and the baculovirus expression vector system (BEVS). |
| spellingShingle | Strain-Damerell, C Mahajan, P Gileadi, O Burgess-Brown, N Medium-throughput production of recombinant human proteins: ligation-independent cloning. |
| title | Medium-throughput production of recombinant human proteins: ligation-independent cloning. |
| title_full | Medium-throughput production of recombinant human proteins: ligation-independent cloning. |
| title_fullStr | Medium-throughput production of recombinant human proteins: ligation-independent cloning. |
| title_full_unstemmed | Medium-throughput production of recombinant human proteins: ligation-independent cloning. |
| title_short | Medium-throughput production of recombinant human proteins: ligation-independent cloning. |
| title_sort | medium throughput production of recombinant human proteins ligation independent cloning |
| work_keys_str_mv | AT straindamerellc mediumthroughputproductionofrecombinanthumanproteinsligationindependentcloning AT mahajanp mediumthroughputproductionofrecombinanthumanproteinsligationindependentcloning AT gileadio mediumthroughputproductionofrecombinanthumanproteinsligationindependentcloning AT burgessbrownn mediumthroughputproductionofrecombinanthumanproteinsligationindependentcloning |