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Nucleobase Protection Strategy for Gene Cloning and Expression
Author(s) -
Kielkowski Pavel,
Brock Nelson L.,
Dickschat Jeroen S.,
Hocek Michal
Publication year - 2013
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201300127
Subject(s) - cloning (programming) , plasmid , restriction enzyme , gene , nucleobase , molecular cloning , cleavage (geology) , microbiology and biotechnology , protecting group , chemistry , dna , biology , synthetic biology , genetics , computational biology , gene expression , paleontology , alkyl , organic chemistry , fracture (geology) , computer science , programming language
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.