TOTAL SYNTHESIS OF A HUMAN-LEUKOCYTE INTERFERON GENE
TOTAL SYNTHESIS OF A HUMAN-LEUKOCYTE INTERFERON GENE
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DOI：
10.1038/292756a0
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发表时间：
1981-01-01
期刊：
影响因子：
64.8
通讯作者：
MARKHAM, AF
中科院分区：
文献类型：
--
作者：
EDGE, MD;GREENE, AR;MARKHAM, AF
完全合成了编码人干扰素-α 1（166个氨基酸残基）的双链DNA的514个碱基对片段，该片段含有起始和终止信号以及用于质粒插入的适当限制酶位点。合成涉及制备66个寡脱氧核糖核苷酸，大小范围从14-21个残基，加上1个脱氧癸核苷酸，通过快速，固相程序和酶连接的寡核苷酸。将合成基因连接到质粒载体上并转化大肠杆菌后，获得含有预期基因序列的克隆。
A 514-base pair fragment of double-stranded DNA coding for human interferon-.alpha.1 (166 amino acid residues), and containing initiation and termination signals plus appropriate restriction enzyme sites for plasmid insertion, was totally synthesized. The synthesis involved preparation of 66 oligodeoxyribonucleotides, ranging in size from 14-21 residues, plus 1 deoxydecanucleotide, by rapid, solid-phase procedures and enzymatic ligation of the oligonucleotides. After ligation of the synthetic gene to a plasmid vector and transformation of Escherichia coli, clones containing the anticipated gene sequence were obtained.