高等学校化学学报

高等学校化学学报

• 研究论文 • 上一篇    下一篇

化学与酶促相结合合成人生长激素基因

赵向阳, 高朝辉, 金璐, 王曦, 孟庆繁, 滕利荣   

  1. 吉林大学生命科学学院, 长春 130012
  • 收稿日期:2008-02-28 修回日期:1900-01-01 出版日期:2008-10-10 发布日期:2008-10-10
  • 通讯作者: 滕利荣

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

ZHAO Xiang-Yang, GAO Chao-Hui, JIN Lu, WANG Xi, MENG Qing-Fan, TENG Li-Rong*   

  1. College of Life Science, Jilin University, Changchun 130012, China
  • Received:2008-02-28 Revised:1900-01-01 Online:2008-10-10 Published:2008-10-10
  • Contact: TENG Li-Rong

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被引次数

1

摘要： 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％.

关键词: 人生长激素, 基因合成, 两步法

Abstract: 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.

Key words: Human growth hormone, Gene synthesis, Two-step method

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