Process for secretory production of a calcium-binding protein
AssigneeChisso Corporation
InventorsShuhei Zenno, Satoshi Inouye
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Abstract
A process for secretory production of a calcium-binding protein very useful for producing heterogenic proteins in Escherichia coli according to gene recombinant technique is provided, which process comprises using Escherichia coli and using a secretory expression system with the promoter of lipoprotein and the gene of signal peptide of outer membrane protein A, according to recombinant DNA technique.
Claims2
A process for producing a fusion protein comprising a calcium-binding protein and biogenic substance of E. coli which facilitates the extracellular secretory production of the fusion protein, which process comprises:
- preparing a recombinant plasmid as a cloning vehicle for expression of the fusion protein in E. coli, said plasmid comprising a first DNA sequence consisting of a lipoprotein promoter region being linked in reading phase with a second DNA sequence located downstream of said first DNA sequence, said second DNA sequence coding for a signal peptide of outer membrane protein A and being linked in reading phase with a third DNA sequence located downstream of said second DNA sequence, said third DNA sequence coding for the fusion protein,
- inserting said plasmid into E. coli to obtain a transformed E. coliand
- cultivating the transformed E. coli in a suitable medium to produce said fusion protein,
- wherein said calcium-binding protein is selected from the group consisting of calmodulin, troponin c, myosin light chain, parvalubumin, vitamin D-dependent calcium-binding proteins, S-100, S-100.beta., calpactin, carpaine/CANP, and oncomodulin.
A process for producing a fusion protein comprising apoaequorin and a biogenic substance in E. coli which facilitates the extracellular secretory production of the fusion protein, which process comprises:
- preparing a recombinant plasmid as a cloning vehicle for expression of the fusion protein in E. coli, said plasmid comprising a first DNA sequence consisting of a lipoprotein promoter region being linked in reading phase with a second DNA sequence located downstream of said first DNA sequence, said second DNA sequence coding for a signal peptide of outer membrane protein A and being linked in reading phase with a third DNA sequence located downstream of said second DNA sequence, said third DNA sequence coding for the fusion protein,
- inserting said plasmid into E. coli to obtain a transformed E. coli, and
- cultivating the transformed E. coli in a suitable medium to produce said fusion protein.
Dependencies are inferred from the claim text.