Christopher L Chio

US Patent:

57211329, Feb 24, 1998

Filed:

May 3, 1995

Appl. No.:

8/434877

Inventors:

Christopher L. Chio - Kalamazoo MI
Rita M. Huff - Kalamazoo MI

Assignee:

The Upjohn Company - Kalamazoo MI

International Classification:

C12N 510
C12N 1512

US Classification:

4352523

Abstract:

A modified gene coding for the human D4 dopamine receptor has been synthesized by chemo-enzymatic methods. The nucleotide sequence of the D4 dopamine receptor gene was changed to reduce the G+C content and to eliminate intronic sequences, while maintaining the published amino acid sequence. Using gene splicing by overlap extension and PCR amplification of long oligonucleotides (>200 bases), 3 synthetic fragments of about 400 base pairs each were amplified, from which the full length gene was assembled. Stable expression of this gene has been achieved in CHO-K1 cells, using an inducible expression system, and in HEK293 cells.

US Patent:

54686156, Nov 21, 1995

Filed:

Jul 1, 1993

Appl. No.:

8/086439

Inventors:

Christopher L. Chio - Kalamazoo MI
Rita M. Huff - Kalamazoo MI

Assignee:

The Upjohn Company - Kalamazoo MI

International Classification:

C12N 1512

US Classification:

435 72

Abstract:

A modified gene coding for the human D4 dopamine receptor has been synthesized by chemo-enzymatic methods. The nucleotide sequence of the D4 dopamine receptor gene was oh:raged to reduce the G+C content and to eliminate intronic sequences, while maintaining the published amino acid sequence. Using gene splicing by overlap extension and PCR amplification of long oligonucleotides (>200 bases), 3 synthetic fragments of about 400 base pairs each were amplified, from which the full length gene was assembled. Stable expression of this gene has been achieved in CHO-K1 cells, using an inducible expression system, and in HEK293 cells.