DNA construct for producing proteinaceous materials in eucaryotic cells

US6455275B1 · kind B1 · utility

Assignee

• The Trustees of Columbia University in the City of New York · US

Inventors

• Richard Axel · New York, US
• Michael Wigler · Lloyd Harbor, US
• Saul J. Silverstein · Dobbs Ferry, US

Key dates

• Filing date: Jun 7, 1995
• Grant date: Sep 24, 2002
• Expiry date: Jun 7, 2015

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12P21/02
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

The invention relates to processes for inserting into eucaryotic cells a multiplicity of DNA molecules which includes genes coding for desired proteinaceous materials. The insertion of multiple copies of desired genes is accomplished by cotransformation with the desired genes and with amplifiable genes for a dominant selectable marker in the presence of successively higher amounts of an inhibitor. Alternatively, the insertion of multiple copies of desired genes is accomplished by transformation using DNA molecules formed by ligating a DNA molecule including the desired gene to a DNA molecule which includes an amplifiable gene coding for a dominant selectable phenotype such as a gene associated with resistance to a drug in the presence of successively higher amounts of an agent such as a drug against which the gene confers resistance so that only those eucaryotic cells into which multiple copies of the amplifiable gene have been inserted survive. Eucaryotic cells into which multiple copies of the amplifiable gene have been inserted additionally include multiple copies of the desired gene and may be used to produce multiple copies of proteinaceous molecules. In this way otherwise rar…