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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.
According to the high expression sequence of Escherichia coli, the nucleotide sequence of the coding region of human interferon α-2b gene was redesigned. A number of complementary DNA fragments were synthesized by chemical synthesis. After the complementary fragments were annealed and ligated, the complete coding region Two clones were respectively cloned into pUC19 plasmid for DNA sequence analysis, and two completely cloned fragments with the correct sequence were selected for recombination, and the human interferon alpha-2b gene containing the complete coding region was obtained which was identical to the designed sequence.