Method for over-expression and rapid purification of biosyntheti

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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435 691, 4353201, 435219, 435226, 435476, 435488, 536 232, 536 234, 530327, 530324, 530329, C12N 1562, C12N 1509

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active

060776943

ABSTRACT:
The subject invention relates to a method of producing and purifying large quantities of a biosynthetic protein.
The gene which codes for the protease is placed between the binding domain of a gene which codes for a binding protein and a gene coding for the target protein of interest. The fused gene construct is inserted in an expression vector which is then introduced into a host cell.

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Louis, John M., Autoprocessing of the HIV-1 Protease Using Purified Wild-Type and Mutated Fusion Proteins Expressed at High Levels in Escherichia coli; Eur. J. Biochem., pp. 199-361-369, 1991.
Bedonelle et al. (1988) Eur. J. Biochem. vol. 171, pp 541-549.
Debouck et al. (1987) PNAS, vol. 84, pp 8903-8906.
S. Scholtissek et al., "A Plasmid Vector System for the Expression of a Triprotein Consisting of .beta.-Galactosidase, a Collagenase Recognition Site and a Foreign Gene Product", Gene 62(1): 55-64, Jan. 1988.
C.V. Maina et al., "An Escherichia coli Vector to Express and purify Foreign Proteins by Fusion to and Separation From Maltose-Binding Protein", Gene 74(2): 365-373, Dec. 1988.
D.B. Smith et al., "Single-Step Purification of Polypeptides Expressed in Escherichia coli as Fusions with Glutathiona S-Transferase", Gene 67(1): 31-40, Jul. 1988.
P.L. Darke et al., "Human Immunodeficiency Virus Protease: Bacterial Expression and Characterizationof the Purified Aspartic Protease", J. Biol. Chem. 264(4): 2307-2312, Feb. 1989.

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