Fusion protein-bound magnetic particles for recombinant producti

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...

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4352523, 4353201, 536 234, 536 237, C12N 1562, C07K 14195

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active

058612858

ABSTRACT:
A fusion DNA sequence, which is obtained by fusing a gene coding for another useful protein to a fragment of a magA gene coding for a protein bound to an organic membrane for covering magnetic particles produced in cells of a magnetic bacterium AMB-1, is expressed in the magnetic bacterium to obtain the protein in a state of being bound to the magnetic particles. According to the present invention, useful proteins such as enzymes and antibodies can be stably obtained in a state of being bound to the organic membrane of the magnetic particles only by cultivating a transformed magnetic bacterium, and separating the magnetic particles produced in cells, without any necessity to perform a treatment such as immobilization. The functional protein immobilized on the magnetic particles can be magnetically controlled. Thus the function can be efficiently exhibited at a desired topical position. Magnetic particles to which a desired protein is bound can be semipermanently produced only by maintaining and cultivating an identical bacterial strain. Since the protein is produced on the magnetic particles, an objective protein can be magnetically separated and recovered in a short period of time. Thus it is possible to perform efficient separation and purification.

REFERENCES:
Matsunaga et al, "Gene Transfer in Magnetic Bacteria: Transposon Mutagenesis and Cloning of Genomic DNA Fragments Required for Masgnetosome Synthesis", Journal of Bacteriology, May 1992, pp. 2748-2753.
Nakamura et al, "Iron-Regulated Expression and Membrane Localization of the MagA Protein in Magnetospirillum sp. Strain AMB-1", J. Biochem.,118, 23-27 (1995).
Nakamura et al, "An Iron-regulated Gene, magA, Encoding an Iron Transport Protein of Magnetospirillumsp. Strain AMB-1", The Journal of Biological Chemistry, vol. 270, No. 47, Nov. 24, 1995, pp. 28395-28396.
Nakamura et al, "Analysis of a Gene, magA, Required for Magnetite Biomineralization in the Bacterium Magnetospirillumsp. AMB-1", Biomineralization 93, 7th International Symposium on Biomineralization, Monaco -17-20 Nov. 1993, Monaco Musee Oceanographique 1995, pp. 121-126.

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