按大肠杆菌高表达序列设计的人干扰素α-2b基因的化学合成和克隆

Design,Chemical Synthesis and Cloning of Human Interferon α-2b Gene According to High-level Expression Sequence of E.Coli

根据大肠杆菌高表达序列重新设计了人干扰素α-2b基因编码区的核苷酸序列,应用化学合成的方法合成了多个互补DNA片段,经互补片段分别退火和连接后,将完整编码区分两段分别克隆于pUC19质粒中,进行DNA序列分析,分别选取序列完全正确的两种克隆片段进行重组.获得了与设计序列完全一致的含有完整编码区的人干扰素α-2b基因.

The coding region′s nucleotide sequence of the human interferon α-2b gene was designed according to high-level expression sequences of E.coli. Several overlapping complementary DNA fragments were synthesized with chemical synthesis method.By annealing and ligating the corresponding oligo-nucleotide,two fragments of 5'end and 3'end that cover the full-length coding region were cloned in plasmid pUC19,respectively.The clones of interest were identified by DNA sequencing.The cloned 5'end and 3'end fragments were recombined into plasmid pUC19.The results showed that the redesigned full-length coding region of human interferon a-2b gene was obtained as expected.