Method of constructing synthetic leader sequences

C - Chemistry – Metallurgy – 12 – N

Patent

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C12N 15/81 (2006.01) C12N 1/19 (2006.01) C12N 15/11 (2006.01) C12N 15/62 (2006.01)

Patent

CA 2098731

2098731 9211378 PCTABS00014 A yeast expression cloning vector comprising the following sequence 5'-SP-Xn-3'-RS-5'-Xm-(NZT)p-Xq-PS-*gene*-3' wherein SP is a DNA sequence encoding a signal peptide, Xn is a DNA sequence encoding n amino acids, wherein n is 0 or an integer of from 1 to about 10 amino acids, RS is a restriction endonuclease recognition site provided at the junction of Xn and Xm, Xm is a DNA sequence encoding m amino acids, wherein m is 0 or an integer from 1 to about 10, (NZT)p is a DNA sequence encoding Asn-Xaa-Thr, wherein p is 0 or 1, Xq is a DNA sequence encoding q amino acids, wherein q is 0 or an integer from 1 to about 10, PS is a DNA sequence encoding a peptide defining a yeast processing site, and *gene* is a DNA sequence encoding a heterologous polypeptide. The vector may be used to construct synthetic leader peptide sequences by inserting random DNA fragments in the "RS" site, culturing a yeast cell transformed with this vector and screening the culture for secretion of the heterologous polypeptide.

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