Open AccessChemical synthesis of genes for human insulin.
Author(s) -
Roberto Crea,
Adam Kraszewski,
Tatsuro Hirose,
Keiichi Itakura
Publication year - 1978
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.75.12.5765
Subject(s) - ecori , human insulin , oligonucleotide , plasmid , gene , bamhi , pbr322 , restriction enzyme , biochemistry , chemistry , biology , base pair , sequence (biology) , combinatorial chemistry , microbiology and biotechnology , insulin , endocrinology
A rapid chemical procedure has been developed and used for the synthesis of 29 oligodeoxyribonucleotides to build synthetic genes for human insulin. The gene for insulin B chain, 104 base pairs, and the one for A chain, 77 base pairs, were designed from the amino acid sequence of human polypeptides. They bear single-stranded cohesive termini for the EcoRI and BamHI restriction endonucleases and are designed to be inserted separately into a pBR322 plasmid. The synthetic fragments, deca- to pentadecanucleotides, were synthesized by a block phosphotriester method with trinucleotides as building blocks. Final purification was by high-performance liquid chromatography. All 29 oligonucleotides were pure and had the correct sequences.