Isolation and characterisation of a gene encoding protein disulphide isomerase, pdiA, from Aspergillus niger
| Autor: | D. J. Jeenes, D. B. Archer, C. Ngiam |
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| Rok vydání: | 1997 |
| Předmět: |
Signal peptide
Protein Folding Transcription Genetic Saccharomyces cerevisiae Molecular Sequence Data Protein Disulfide-Isomerases Biology Protein Sorting Signals Polymerase Chain Reaction Gene Expression Regulation Enzymologic chemistry.chemical_compound Gene Expression Regulation Fungal Genetics Escherichia coli Amino Acid Sequence Cloning Molecular Isomerases Promoter Regions Genetic Gene DNA Primers Base Sequence Sequence Homology Amino Acid Endoplasmic reticulum Tunicamycin General Medicine biology.organism_classification Blotting Northern Introns Anti-Bacterial Agents Blotting Southern Kinetics Biochemistry chemistry Foldase Protein folding Heterologous expression Aspergillus niger Oxidoreductases Molecular Chaperones |
| Zdroj: | Current genetics. 31(2) |
| ISSN: | 0172-8083 |
| Popis: | Current strategies to improve the secretion of heterologous proteins from Aspergillus niger include the manipulation of chaperones and foldases specific to the endoplasmic reticulum (ER). Here we report the isolation of a gene, pdiA, encoding a putative protein disulphide isomerase (PDI) from A. niger using the Saccharomyces cerevisiae PDI gene as a probe. Sequencing of a genomic clone and RT-PCR products predict a 515-aa protein comprising a 20-aa ER-translocation signal sequence and a 495-aa mature protein (Mr = 54.3 kDa). The predicted protein also contains two thiol oxidoreductase active sites with a -CGHC- motif and a carboxy terminal -HDEL ER-retention signal. Three introns were identified within the pdiA gene and Southern- and dot-blot analysis indicates that the gene is present in a single copy. Northern-blot analysis shows a transcript of the predicted size. Sequence homology to a motif associated with protein trafficking and the induction of chaperones has been identified in the pdiA promoter. Transcription of pdiA is induced 3–4-fold after treatment with tunicamycin, an inhibitor of N-linked glycosylation. The kinetics of induction suggest that pdiA expression is not part of the primary stress response. |
| Databáze: | OpenAIRE |
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