Chemical synthesis and cloning of a gene for human β-urogastrone

摘要

A DNA duplex coding for the 53 amino acids of human β-urogastrone has been synthesised. Computer assisted design of the gene included restriction endonuclease sites for plasmid insertion, a termination codon and two triplets coding for lysine at the 5′-end of the structural gene. The synthesis involved preparation of 23 oligodeoxyribo-nucleotides by phosphotriester procedures coupled to rapid HPLC techniques. The gene was constructed in two halves by enzymatic ligation of the oligonucleotides and cloned into a specially constructed chimeric plasmid vector. Escherichia coli K12 MRC8 was transformed by the plasmid and clones containing the full gene sequence were isolated and characterised.