Highly efficient protein trans-splicing by a naturally split DnaE intein from Nostoc punctiforme |
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Authors: | Iwai Hideo Züger Sara Jin Jennifer Tam Pui-Hang |
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Affiliation: | Department of Chemistry, 110 Science Place, Saskatoon, SK, Canada, S7N 5C9. hideo.iwai@helsinki.fi |
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Abstract: | Protein trans-splicing by the naturally split intein of the gene dnaE from Nostoc punctiforme (Npu DnaE) was demonstrated here with non-native exteins in Escherichia coli. Npu DnaE possesses robust trans-splicing activity with an efficiency of > 98%, which is superior to that of the DnaE intein from Synechocystis sp. strain PCC6803 (Ssp DnaE). Both the N- and C-terminal parts of the split Npu DnaE intein can be substituted with the corresponding fragment of Ssp DnaE without loss of trans-splicing activity. Protein splicing with the Npu DnaEN is also more tolerant of amino acid substitutions in the C-terminal extein sequence. |
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Keywords: | IPTG, isopropyl β- smallcaps" >d-thiogalactoside SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis MALDI-TOF, matrix-assisted laser desorption and ionization-time-of-flight GB1, the B1 domain of IgG binding protein, protein G |
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