Construction of three new Gateway® expression plasmids for Trypanosoma cruzi
We present here three expression plasmids for Trypanosoma cruzi adapted to the Gateway® recombination cloning system. Two of these plasmids were designed to express trypanosomal proteins fused to a double tag for tandem affinity purification (TAPtag). The TAPta...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Fundação Oswaldo Cruz (FIOCRUZ)
2014-12-01
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| Series: | Memorias do Instituto Oswaldo Cruz |
| Subjects: | |
| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762014000801081&lng=en&tlng=en |
| _version_ | 1797721263792193536 |
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| author | Victoria L Alonso Carla Ritagliati Pamela Cribb Esteban C Serra |
| author_facet | Victoria L Alonso Carla Ritagliati Pamela Cribb Esteban C Serra |
| author_sort | Victoria L Alonso |
| collection | DOAJ |
| description | We present here three expression plasmids for Trypanosoma cruzi adapted to the Gateway® recombination cloning system. Two of these plasmids were designed to express trypanosomal proteins fused to a double tag for tandem affinity purification (TAPtag). The TAPtag and Gateway® cassette were introduced into an episomal (pTEX) and an integrative (pTREX) plasmid. Both plasmids were assayed by introducing green fluorescent protein (GFP) by recombination and the integrity of the double-tagged protein was determined by western blotting and immunofluorescence microscopy. The third Gateway adapted vector assayed was the inducible pTcINDEX. When tested with GFP, pTcINDEX-GW showed a good response to tetracycline, being less leaky than its precursor (pTcINDEX). |
| first_indexed | 2024-03-12T09:31:58Z |
| format | Article |
| id | doaj.art-33a8aa1579f744a3bf8f773e51ee8420 |
| institution | Directory Open Access Journal |
| issn | 1678-8060 |
| language | English |
| last_indexed | 2024-03-12T09:31:58Z |
| publishDate | 2014-12-01 |
| publisher | Fundação Oswaldo Cruz (FIOCRUZ) |
| record_format | Article |
| series | Memorias do Instituto Oswaldo Cruz |
| spelling | doaj.art-33a8aa1579f744a3bf8f773e51ee84202023-09-02T13:54:07ZengFundação Oswaldo Cruz (FIOCRUZ)Memorias do Instituto Oswaldo Cruz1678-80602014-12-0110981081108510.1590/0074-0276140238S0074-02762014000801081Construction of three new Gateway® expression plasmids for Trypanosoma cruziVictoria L AlonsoCarla RitagliatiPamela CribbEsteban C SerraWe present here three expression plasmids for Trypanosoma cruzi adapted to the Gateway® recombination cloning system. Two of these plasmids were designed to express trypanosomal proteins fused to a double tag for tandem affinity purification (TAPtag). The TAPtag and Gateway® cassette were introduced into an episomal (pTEX) and an integrative (pTREX) plasmid. Both plasmids were assayed by introducing green fluorescent protein (GFP) by recombination and the integrity of the double-tagged protein was determined by western blotting and immunofluorescence microscopy. The third Gateway adapted vector assayed was the inducible pTcINDEX. When tested with GFP, pTcINDEX-GW showed a good response to tetracycline, being less leaky than its precursor (pTcINDEX).http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762014000801081&lng=en&tlng=enTrypanosoma cruziexpression vectors - Gateway® systemtandem affinity purificationpTcINDEX |
| spellingShingle | Victoria L Alonso Carla Ritagliati Pamela Cribb Esteban C Serra Construction of three new Gateway® expression plasmids for Trypanosoma cruzi Memorias do Instituto Oswaldo Cruz Trypanosoma cruzi expression vectors - Gateway® system tandem affinity purification pTcINDEX |
| title | Construction of three new Gateway® expression plasmids for Trypanosoma cruzi |
| title_full | Construction of three new Gateway® expression plasmids for Trypanosoma cruzi |
| title_fullStr | Construction of three new Gateway® expression plasmids for Trypanosoma cruzi |
| title_full_unstemmed | Construction of three new Gateway® expression plasmids for Trypanosoma cruzi |
| title_short | Construction of three new Gateway® expression plasmids for Trypanosoma cruzi |
| title_sort | construction of three new gateway 174 expression plasmids for trypanosoma cruzi |
| topic | Trypanosoma cruzi expression vectors - Gateway® system tandem affinity purification pTcINDEX |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762014000801081&lng=en&tlng=en |
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