Synthesis of the leu-enkephalin gene

The synthesis of Leu-enkephalin gene by an alternative approach was described in this paper. The sequence of the synthetic gene, 26 base-pairs in length, was derived from the amino acid sequence of the hormone peptide Leu-enkephalin. It bears single-stranded cohesive termini for the restriction endonucleases EcoR I and BamH I so that it may be inserted into a pBR 322 plasmid. The 4 deoxyoligonucleotide fragments, varying in length from octanucleotide to octadecanucleotide, were synthesized by an improved phosphotriester method developed in our laboratory. All chemically synthetic fragments were pure and had the correct sequences. The assembly of the Leu-enkephalin gene was carried out in a one-step ligation reaction catalysed by T4-DNA ligase with good yield. Finally, the purified products from polyacrylamide gel electrophoresis had the correct joining-points as predicted.