High-Level Direct Expression of Semi-Synthetic Human Interleukin-6 in Escherichia coli and Production of N-Terminus Met-Free Product
High-Level Direct Expression of Semi-Synthetic Human Interleukin-6 in Escherichia coli and Production of N-Terminus Met-Free Product
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半合成人白细胞介素 6 在大肠杆菌中的高水平直接表达及 N 端无 Met 产品的生产
DOI：
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发表时间：
1990
期刊：
影响因子：
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通讯作者：
Kazuhiko Yamada
中科院分区：
文献类型：
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作者：
H. Yasueda;K. Nagase;A. Hosoda;Y. Akiyama;Kazuhiko Yamada
我们建立了一种在大肠杆菌中高效表达重组人白细胞介素6(rhIL-6)的系统。在该系统中，(I)hIL-6基因的自然N-末端编码区被含有富含A-T的密码子的合成序列取代，(Ii)采用双Shine-Dalgarno(SD)序列，(Iii)在起始密码子的前面插入一个富含A-T的片段，以避免该区域可能存在的mRNA二级结构，以及(Iv)hIL-6基因的天然琥珀终止密码子被改变为鸵鸟终止密码子。高水平合成的hll-6多肽形成胞质内包涵体。复性后，氨基肽酶-P在体外将N端的蛋氨酸去掉。纯化的重组人IL-6具有与人T细胞培养中自然产生的IL-6相当的B细胞分化活性。
We have developed a direct expression system for high-level production of recombinant human interleukin-6 (rhIL-6) in Escherichia coli. In this system, (i) the natural N-terminal coding region of the hIL-6 gene was replaced by a synthetic sequence containing A-T rich codons, (ii) dual Shine-Dalgarno (SD) sequences were employed, (iii) an A-T rich segment was inserted in front of the initiation codon to avoid putative mRNA secondary structure in the region and (iv) the natural amber termination codon of the hIL-6 gene was changed to an ocher stop codon. The hlL-6 polypeptide, synthesized at a high level, formed cytoplasmic inclusion bodies. After refolding, the N-terminal methionine was removed by aminopeptidase-P in vitro. The purified recombinant hIL-6 had B-cell differentiation activity equivalent to natural IL-6 from a human T-cell culture.
高水平表达生物工程、不含半胱氨酸的肝细胞刺激因子（白细胞介素 6）样蛋白。
DOI：
10.1073/pnas.85.24.9426
发表时间：
1988
影响因子：
11.1
作者：
Jambou,RC;Snouwaert,JN;Bishop,GA;Stebbins,JR;Frelinger,JA;Fowlkes,DM
通讯作者：
Fowlkes,DM