单链噬菌体载体中真核基因的克隆：人类干扰素基因

Cloning of eukaryotic genes in single-strand phage vectors: the human interferon genes.

作者信息

Bowden D W, Mao J, Gill T, Hsiao K, Lillquist J S, Testa D, Vovis G F

出版信息

Gene. 1984 Jan;27(1):87-99. doi: 10.1016/0378-1119(84)90241-5.

Abstract

Using oligonucleotide probes with defined sequences, we have selected clones from a human lymphocyte cDNA library which represent human leukocyte (HuIFN-alpha) and fibroblast (HuIFN-beta) interferon gene sequences. Double-stranded f1 phage DNA was used as the vector for initial cloning of cDNA. Clones carrying interferon gene sequences were identified by hybridization with the oligonucleotide probes. The same oligonucleotide probes were used as primers for dideoxy chain termination sequencing of the clones. One HuIFN-alpha clone, 201, has a nucleotide sequence different from published HuIFN-alpha sequences. Under control of the lacUV5 promoter, the 201 gene has been used to express biologically active HuIFN-alpha in Escherichia coli.

摘要

我们使用具有特定序列的寡核苷酸探针，从人淋巴细胞cDNA文库中筛选出了代表人类白细胞（HuIFN-α）和成纤维细胞（HuIFN-β）干扰素基因序列的克隆。双链f1噬菌体DNA被用作cDNA初始克隆的载体。携带干扰素基因序列的克隆通过与寡核苷酸探针杂交来鉴定。同样的寡核苷酸探针被用作这些克隆进行双脱氧链终止测序的引物。一个HuIFN-α克隆，即201，其核苷酸序列与已发表的HuIFN-α序列不同。在lacUV5启动子的控制下，201基因已被用于在大肠杆菌中表达具有生物活性的HuIFN-α。