Synthesis of Codon-optimized Human Interleukin-18 Gene by Combination of Chemical and Enzymatic Method

摘要

According to the amino acid sequence and codon preference of E. coli,the human interleukin-18(IL-18) gene was optimized to avoid the rare codons. The total length of the synthesized gene is 571 bp; 18 oligonucleotides,DNA fragments were designed and synthesized by the phosphoramidite four-step chemical method. The whole DNA sequence was synthesized by a one-step total gene synthesis method,and then inserted in pUC18 vector. Five positive clones identified by blue-white colony screening were sent to Shanghai Sangon Biological Engineering Technology and Service Co.,Ltd. for sequencing. The sequencing result shows that one clone contained the complete correct gene in all the five positive clones.