Expression of a biologically active fragment of human IgE epsilon chain in Escherichia coli.
Expression of a biologically active fragment of human IgE epsilon chain in Escherichia coli.
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人 IgE ε 链的生物活性片段在大肠杆菌中的表达。
DOI：
10.1073/pnas.81.17.5369
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发表时间：
1984
影响因子：
11.1
通讯作者：
Ishizaka,T
中科院分区：
文献类型：
--
作者：
Liu,FT;Albrandt,KA;Bry,CG;Ishizaka,T
从人IgE分泌型骨髓瘤U266细胞中克隆了相应于人IgE重链mRNA的cDNA。部分核苷酸序列分析表明，克隆的cDNA含有约三分之二的CH 2和所有的CH 3和CH 4结构域的编码区以及3 '非翻译区。将此cDNA插入表达载体pUC 7中，用125I标记的山羊抗人IgE作为探针，通过蛋白质印迹分析证明ε链片段在大肠杆菌中表达。表达产物在山羊抗人IgE缀合的Sepharose 4B柱上纯化，发现多肽保留与人嗜碱性粒细胞的结合活性。
cDNA corresponding to human IgE heavy (epsilon) chain mRNA was cloned from human IgE-secreting myeloma U266 cells. Partial nucleotide sequence analysis demonstrated that the cloned cDNA contained the coding region for about two-thirds of the CH2 and all of the CH3 and CH4 domains as well as the 3'-untranslated region. This epsilon cDNA was inserted into expression vector pUC7 and expression of an epsilon-chain fragment in Escherichia coli was demonstrated by protein blot analysis using 125I-labeled goat anti-human IgE as probe. The expression product was purified on a column of goat anti-human IgE-conjugated Sepharose 4B and the polypeptide was found to retain binding activity to human basophils.