Chemical synthesis of a biologically active gene for human immune interferon-gamma. Prospect for site-specific mutagenesis and structure-function studies.
A 453-base pair DNA duplex consisting of a gene coding for human interferon-gamma and initiation and termination signals plus appropriate restriction enzyme sites for plasmid insertion has been totally synthesized. The synthesis involved preparation of 66 oligodeoxynucleotides by a modified, solid phase phosphite procedure and enzymatic ligation of the oligonucleotides. The gene, when inserted into a previously constructed expression vector, was expressed in Escherichia coli, demonstrating functional activity for the synthetic gene. Several strategically located restriction cleavage sites have been introduced into the sequence. This provides a convenient system for site-specific mutagenesis for structure-function studies.
D000595:Amino Acid Sequence; D001482:Base Composition; D001483:Base Sequence; D004262:DNA Restriction Enzymes; D005796:Genes; D005813:Genes, Synthetic; D006801:Humans; D007371:Interferon-gamma / Q000235:genetics*; D009154:Mutation; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis; D010957:Plasmids; D013329:Structure-Activity Relationship
D009838:Oligodeoxyribonucleotides; D007371:Interferon-gamma; D004262:DNA Restriction Enzymes