Nucleobase Protection Strategy for Gene Cloning and Expression

Abstract

Protecting group chemistry meets molecular biology: Chemically modified dATP carrying a bulky triethylsilylethynyl group was used in a PCR-based synthesis of a gene internally protected against cleavage by restriction endonucleases. The unmodified flanking regions were cleaved for cloning into a plasmid which was replicated by E. coli, and used for protein production.

Keywords

• DNA
• polymerase chain reaction
• protecting groups
• protein production
• restriction endonucleases