Trans-dominant suppressor genes for oligomeric proteins

Details Trans-dominant suppressor genes for oligomeric proteins Trans-dominant suppressor genes for oligomeric proteins

C - Chemistry, Metallurgy

12

N

C12N 15/10 (2006.01) A01K 67/027 (2006.01) C07K 14/49 (2006.01) C12N 15/12 (2006.01) C12N 15/18 (2006.01) C12N 15/85 (2006.01) A61K 38/00 (2006.01)

Patent

CA 2083164

2083164 9118001 PCTABS00008 A method of making trans-dominant, suppressor genes encoding mutant subunits of two different categories of oligomeric proteins is disclosed. In one category, the protein is composed of subunits at least one of which is initially synthesized containing extraneous peptide material. This extraneous material is subsequently removed during normal processing by cleavage at a proteolytic cleavage site to generate the mature form of the subunit. According to the invention, single stranded DNA encoding the initial form of the indicated subunit is cloned, and the base sequence of the DNA in the region encoding the proteolytic cleavage site is modified by site directed mutagenesis so that processing of the translation product of the gene to the mature form of the subunit is prevented. In the other category, the oligomeric protein is one in which the monomers are bonded together by at least two disulfide bonds, at least one of which is essential for the catalytic activity of the protein. In the method of the invention, single stranded DNA encoding one of the subunits is cloned, and the base sequence of the DNA in the region encoding a cysteine residue essential for catalytic activity is modified by site directed mutagenesis.

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