GENE CODING MUTANT-TYPE HUMAN LYSOZYME

摘要

PURPOSE:To obtain the new gene coding a polypeptide with cysteine at specific sites along the amino acid sequence of a natural-type human lysozyme substituted with alanine, capable of efficiently secrete human lysozyme through manifestation in host cells. CONSTITUTION:A plasmid pERI8602 containing a gene coding a natural-type human lysozyme is subjected to restriction enzyme treatment to cut out a XhoI- SmaI restriction fragment containing a signal peptide and a nucleotide sequence coding a human lysozyme, and this fragment is linked to a phage DNA followed by mixing with an oligonucleotide coding a short sequence with cysteine at the 77th site and 95th site along the amino acid sequence of the natural-type human lysozyme substituted with alanine, followed by annealing and ligation, thus obtaining the objective gene coding mutant-type human lysozyme coding a polypeptide with cysteine at the 77th site and 95th site along the amino acid sequence of the natural-type human lysozyme substituted with alanine.