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 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).
| Main Authors: | , , , |
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| Format: | Digital revista |
| Language: | English |
| Published: |
Instituto Oswaldo Cruz, Ministério da Saúde
2014
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| Online Access: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762014000801081 |
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