化学与酶促相结合合成人生长激素基因

根据人生长激素(hGH)的氨基酸序列和大肠杆菌密码子的偏好性, 全面优化hGH的密码子, 添加表达调控元件后合成序列的全长为1040 bp, 采用化学方法拆分成30条单链寡核苷酸. 采用改进的两步法拼接成全基因, 得到2个全序列正确的基因. DA-PCR和OE-PCR拼接产物经T7核酸内切酶Ⅰ处理, 合成基因中的碱基错误率降低了93.93％.

Synthesis of Human Growth Hormone Gene by the Combination of Chemical and Enzymatic Method

According to the amino acid sequence and codon preference of E.coli,the human growth hormone(hGH) gene was optimized to avoid rare codons.The total length of the synthesized gene was 1040 bp because a promoter and a signal peptide sequence were added into it.Thirty oligonucleotides were designed and synthesized.The whole DNA sequence was synthesized by the improved two-step method.The sequencing result show that two complete correct gene were synthesised.The error rate of synthesized gene treated by T7 endonuclease I reduced 93.93% in comparison with the standard two-step synthesis method.