Patent ID: 6573100
Filing Date: 2003-06-03
Classification: A61K,A61P,C07K,C12N

Abstract:
A method for making a plasmid comprising a heterologous nucleic acid molecule encoding a product, comprising:(i) introducing a heterologous nucleic acid molecule encoding a product operatively linked to a promoter into a plasmid, said plasmid further comprising a DNA sequence encoding an antibiotic resistance selection marker operatively linked to a promoter; (ii) amplifying said plasmid; (iii) cleaving said amplified plasmid to form two linear fragments, the first of which comprises said heterologous nucleic acid molecule operatively linked to a promoter, the second of which comprises said DNA sequence molecule encoding said selection marker operatively linked to a promoter; (iv) circularizing each of said fragments to form two plasmids, the first of which comprises said heterologous nucleic acid molecule operatively linked to a promoter and lacking said selection marker, the second of which comprises said DNA sequence encoding said antibiotic resistance selection marker operatively linked to a promoter and lacking said heterologous nucleic acid molecule; (v) separating the plasmids obtained in (iv); and (vi) isolating the plasmid comprising the heterologous nucleic acid molecule encoding a product operatively linked to a promoter.