Efficient expression in Escherichia coli of two species of human interferon-alpha and their hybrid molecules.
Efficient expression in Escherichia coli of two species of human interferon-alpha and their hybrid molecules.
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两种人干扰素-α及其杂合分子在大肠杆菌中的高效表达。
DOI：
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发表时间：
1985
期刊：
影响因子：
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通讯作者：
N. Raj
中科院分区：
文献类型：
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作者：
J. Mizoguchi;P. Pitha;N. Raj
从仙台病毒感染的Namalva细胞构建的cDNA文库中鉴定出一种新的干扰素(IFN)- α cDNA （IFN- α I’）。除了编码区有两个核苷酸外，该cDNA的核苷酸序列与先前鉴定的ifn - α 1基因具有同源性（Lawn et al., 1981）。利用携带lpp/lac启动子和“失控”复制子的载体构建了一个指导成熟ifn - α I′肽合成的表达质粒。此外，将ifn - α I'和ifn - α A在位于121-122个氨基酸位置的共同限制性内切酶位点进行体外重组，构建杂交基因。虽然IFN α A和IFN α I′对人类细胞的特异性抗病毒和抗细胞活性相当，但IFN α I′对小鼠细胞的抗病毒活性比IFN α A高125倍。杂交IFN对人类和牛细胞的特异性抗病毒活性与氨基末端亲本IFN肽相似。anticellular活动在人类细胞的α/αα的混合是高的,我/α混合低于父母IFN-alpha IFN-alpha我”。
A new type of interferon (IFN)-alpha cDNA (IFN-alpha I') was identified in a cDNA library constructed from Namalva cells infected with Sendai virus. The nucleotide sequence of this cDNA showed homology, with the exception of two nucleotides in the coding region, with the previously identified IFN-alpha I gene (Lawn et al., 1981). An expression plasmid which directs the synthesis of the mature IFN-alpha I' peptide was constructed using vectors carrying the lpp/lac promoter and "runaway" replicon. Furthermore, hybrid genes were constructed by in vitro recombination of IFN-alpha I' and IFN-alpha A at a common restriction endonuclease site located at amino acid positions 121-122. While the specific antiviral and anticellular activities of IFN alpha A and IFN-alpha I' on human cells were comparable, the antiviral activity of IFN-alpha I' on mouse cells was 125-fold higher than that of IFN-alpha A. The specific antiviral activities of the hybrid IFNs on human and bovine cells were similar to that of the amino-terminal parental IFN peptide, while the anticellular activities on human cells of the alpha A/alpha I' hybrid were higher and that of the alpha I'/alpha A hybrid were lower than the parental IFN-alpha A and IFN-alpha I'.
使用大肠杆菌脂蛋白基因构建多功能表达克隆载体。
DOI：
10.1002/j.1460-2075.1982.tb01244.x
发表时间：
1982
期刊：
The EMBO journal
影响因子：
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作者：
Nakamura,K;Inouye,M
通讯作者：
Inouye,M