Cloning and expression in Escherichia coli of the gene for mouse tumor necrosis factor.
Cloning and expression in Escherichia coli of the gene for mouse tumor necrosis factor.
复制标题
小鼠肿瘤坏死因子基因的克隆及其在大肠杆菌中的表达。
DOI：
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发表时间：
1988
期刊：
影响因子：
--
通讯作者：
Hirataka Ito
中科院分区：
文献类型：
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作者：
Takashi Shirai;Noriko Shimizu;S. Shiojiri;Sadayuki Horiguchi;Hirataka Ito
小鼠肿瘤坏死因子（TNF）基因是从小鼠基因组文库中分离出来的。使用具有改进的引物延伸反应条件的自动DNA测序仪以及标准放射性同位素方法测定了该基因的整个序列。将该基因的核苷酸序列与小鼠TNF cDNA的核苷酸序列进行比较表明，小鼠基因由四个外显子组成，与兔和人TNF基因一样。小鼠、兔和人TNF基因的5'侧翼区域具有很强的核苷酸序列同源性，表明调节TNF基因表达的机制是高度保守的。使用定点诱变构建的质粒实现成熟小鼠 TNF 的直接表达。大肠杆菌产生的纯化小鼠 TNF 对小鼠 L 细胞具有细胞毒性。
The mouse tumor necrosis factor (TNF) gene was isolated from a mouse genomic library. The entire sequence of the gene was determined using both an automated DNA sequencer with improved primer extension reaction conditions as well as the standard radioisotopic method. Comparison of the nucleotide sequence of the gene with that of mouse TNF cDNA showed that the mouse gene consists of four exons, like rabbit and human TNF genes. There is strong nucleotide sequence homology in the 5'-flanking region among the mouse, rabbit, and human TNF genes, suggesting that the mechanisms regulating TNF gene expression are highly conserved. Direct expression of mature mouse TNF was achieved using a plasmid constructed by site-directed mutagenesis. Purified mouse TNF produced in Escherichia coli showed cytotoxicity to mouse L cells.
DOI：
10.1073/pnas.83.6.1670
发表时间：
1986-03-01
影响因子：
11.1
作者：
CAPUT, D;BEUTLER, B;CERAMI, A
通讯作者：
CERAMI, A
影响因子：
3.5
作者：
HENIKOFF, S
通讯作者：
HENIKOFF, S