title: 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.
authors.affiliation: Institute of Organic Chemistry and Biochemistry, Academy of Science of the Czech Republic, Gilead Sciences & IOCB Research Center, Flemingovo nám. 2, 16610 Prague 6, Czech Republic.
authors.forename: Pavel
authors.initials: P
authors.lastname: Kielkowski
authors.forename: Nelson L
authors.initials: NL
authors.lastname: Brock
authors.forename: Jeroen S
authors.initials: JS
authors.lastname: Dickschat
authors.forename: Michal
authors.initials: M
authors.lastname: Hocek
mesh_terms: D003001:Cloning, Molecular / Q000379:methods*; D003838:Deoxyadenine Nucleotides / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction
chemical_list: D003838:Deoxyadenine Nucleotides; C026600:2'-deoxyadenosine triphosphate