Synthesis and cloning of the Human Lysozyme gene
人溶菌酶基因的合成和克隆

A Human lysozyme gene has been chemically synthesized by solid-phase phosphoramidite method. The synthetic gene consisted of 409 bp contains the structural gene coding for human lysozyme, a start signal ATG and BamH I restriction site at 5' end, and two stop signal TAA TGA and Sph I restriction site at 3'. Whole gene was divided into 24 fragments with chain lengths of 26-38, which was prepared respectively. The oligonucleotides were joined to form DNA duplexes, human lysozyme gene, by two methods of ligation. The synthetic gene was cloned into vector M13. The positive colonies were confirmed by dot-blot hybridization and analysis by restriction enzymes. The DNA sequence of the cloned human lysozyme gene was proved to be correct by M13 dideoxynucleotide chain termination method.