Expression and secretion of recombinant actinidin (cysteine protease) in Saccharomyces cerevisiae
Two variants of actinidin-encoding DNA sequences have been constructed and expressed in yeast Saccharomyces cerevisiae. The two variants represented different regions of full-length actinidin cDNA. The first variant (R2) consisted of mature actinidin sequences (starting from amino acid number 127) p...
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| Format: | Article NonPeerReviewed |
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[Yogyakarta] : Universitas Gadjah Mada
2003
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| Online Access: | https://repository.ugm.ac.id/23761/ http://i-lib.ugm.ac.id/jurnal/download.php?dataId=6721 |
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| Summary: | Two variants of actinidin-encoding DNA sequences have been constructed and expressed in yeast Saccharomyces cerevisiae. The two variants represented different regions of full-length actinidin cDNA. The first variant (R2) consisted of mature actinidin sequences (starting from amino acid number 127) plus the C-terminal extension up to amino acids number 384. The second variant (R5) consisted of the N-terminal extension, starting from amino acid number 34, spanned through the mature actinidin actinidin sequences, plus the whole C-terminal extension (amino acid number 389). It was observed that, upon expression in S, cerevisiae using the yeast expression-secretion vector, only the R2 variant resulted in the secreted protein reacted immunogenically with anti-actinidin antibody. The secreted protein, however, had a lower molecular weight as compared to the native actinidin, suggesting that the precursor actinidin was not correctly processed. Nevertheles, it was demonstrated that removal of specific amino acid motifs within the actinidin precursor had resulted in secretion of actinidin into the culture supernatant.
Keywords: actinidin -Saccharomyces cerevisiae -protein translocation - protein engineering |
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