Expression in mammalian cells of a gene fromStreptomyces albonigerconferring puromycin resistance

Key Points

• Determine whether the puromycin N-acetyl transferase gene from Streptomyces alboniger can be expressed in mammalian cells to confer resistance to puromycin.
• Cloned the Streptomyces alboniger puromycin N-acetyl transferase gene adjacent to the SV40 early promoter within a mammalian–Escherichia coli shuttle vector.
• Introduced the recombinant vector into mammalian VERO cells and assayed for enzymatic activity and cell survival under puromycin selection.
• Transfected VERO cells demonstrated functional expression of puromycin N-acetyl transferase enzymatic activity.
• The cloned gene operated successfully as a dominant marker, permitting transformed mammalian cells to grow in the presence of puromycin.