RANDOM CODON BASED MUTAGENESIS USING TRANSPOSON

摘要

PURPOSE: Provided is random codon based mutagenesis using transposon for directed evolution of the protein, thereby providing mutant protein having improved properties to be directed and polynucleotide encoding the same. CONSTITUTION: A directed evolution method of polypeptide and its coding polynucleotide comprises the steps of: preparing a target double stranded DNA in which transposon is deleted at random position; deleting the duplicated nucleotide sequence of the target DNA with transposon derived nucleotide sequence; inserting codon nucleotide into the region where duplicated nucleotide sequence is deleted, and deleting transposon derived nucleotide sequence and consecutive 3 bp nucleotide sequence of the target DNA; self-ligation of two ends of the target DNA to manufacture mutant library; and expressing the mutant library in an appropriate host cell and screening a target protein to give mutant polypeptide having directed properties or mutant polynucleotide encoding the same.