Process for producing apoaequorin

Details Process for producing apoaequorin Process for producing apoaequorin

1988-11-18

1992-08-18

Russel, Jeffrey E.

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Recombinant dna technique included in method of making a...

530350, 530416, 530419, 530420, C07K 322, C07K 324, C07K 1508, C12P 2100

Patent

active

051399377

ABSTRACT:
A process for producing the protein part of Ca.sup.+2 -binding photoprotein aequorin (apoaequorin) according to a recombinant DNA technique, and a process for purifying the resulting apoaequorin are provided, which production process comprises cultivating a strain having an expression vector piP-HE outside the bacterial bodies transformed into Escherichia coli, followed by separating the resulting culture solution into the bacterial bodies and a culture filtrate, and recovering the culture filtrate, and which purification process comprises adding an acid to the culture filtrate so as to give a pH of 4.7 or less, followed by recovering the resulting white precipitates, dissolving the white precipitates in a buffer solution, reducing the solution, subjecting the resulting apoaequorin fraction to adsorption treatment according to anion exchange chromatography and subjecting the resulting apoaequorin to gel filtration.

REFERENCES:
patent: 4643969 (1987-02-01), Inouye et al.
Ghrayeb et al., "Secretion cloning vectors in Escherichia coli", EMBO Journal 3:2437-42 1984.
Prasher et al., "Cloning and Expression of the cDNA Coding for Aequorin . . . ", Biochem. Biophys. Res. Comm. 126:1259-68, 1985.
Tsuji et al., "Site-specific mutagenesis of the calcium-binding photoprotein aequorin," PNAS 83:8107-11, Nov. 1986.
Bjurstrom, "Biotechnology", Chemical Engineering Feb. 18, 1985, pp. 126, 142-143, 151-154.
Inouye et al., "Cloning and sequence analysis of cDNA for the luminescent protein aequorin," PNAS vol. 82, pp. 3154-3158, May 1985.
Johnson et al., "Introduction to the Bioluminescence of Medusae . . . ", Methods in Enzymology LVII, 1978, pp. 271, 287-290.
Prasher et al., "Isolation and Expression of a cDNA Coding for Aequorin . . . ", Methods in Enzymology vol. 133, 1986, pp. 288-298.

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