Plasmid for expressing phosphorylated recombinant proteins in a

Details Plasmid for expressing phosphorylated recombinant proteins in a Plasmid for expressing phosphorylated recombinant proteins in a

1995-11-06

1998-01-20

Elliott, George C.

Chemistry: molecular biology and microbiology

Vector, per se

435 691, 4351723, 435194, 530360, C12N 1563, C12N 1572, C12N 1573, C12N 912

Patent

active

057100446

ABSTRACT:
The present invention provides a plasmid containing a promoter sequence, a nucleotide sequence encoding an exogenous protein, and a nucleotide sequence encoding an enzyme capable of modifying the exogenous protein. In a specific embodiment of the invention the encoded exogenous protein is human .beta.-casein and the encoded enzyme is a human kinase capable of phosphorylating recombinant .beta.-casein in a bacterial system. Phosphorylated recombinant human .beta.-casein having 3 or more phosphate groups and synthesized using the plasmid of the invention is shown to have the same bioactivity as native human .beta.-casein.

REFERENCES:
patent: 5459051 (1995-10-01), Macarenhas et al.
patent: 5460950 (1995-10-01), Barr et al.
Jeng et al., Biochem. Biophys. Res. Comm. 203(1):225-230 (1994).
Shi et al., Proc. Natl. Acad. Sci. USA 91:2767-2771 (1994).
Hansson et al., Prot. Exp. and Purif. 4:373-381 (1993).
Chemical Abstracts, vol. 112, No. 19, 7 May 1990, p. 221 Jimenez-Flores, R. et al., "Expression of bovine .beta.-casein in Saccharomyces cerevisiae and characterization of the protein produced in vivo."
309 Journal of Dairy Science (ADSA), 71, (1988) Oct., No. 10, Champaign, IL, USA. Jimeneze-Flores, R. et al., "Genetic engineering of the caseins to modify the behavior of milk during processing : A review".
Chemical Abstracts, vol. 120, No. 3, 17 Jan. 1994, Columbus, OH, US, Abstract 25819, Hansson, Lennart et al., "Expression of human milk.beta.-casein in Escherichia coli . . . ".

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