RECOMBINANT HUMAN HEMOGLOBIN - EXPRESSION AND REFOLDING OF BETA-GLOBIN FROM ESCHERICHIA-COLI

被引：33

作者：

FRONTICELLI, C ^{[1
]}

ODONNELL, JK ^{[1
]}

BRINIGAR, WS ^{[1
]}

机构：

[1] TEMPLE UNIV,DEPT CHEM,PHILADELPHIA,PA 19122

来源：

JOURNAL OF PROTEIN CHEMISTRY | 1991年 / 10卷 / 05期

关键词：

HEMOGLOBIN; RECOMBINANT HEMOGLOBIN; PROTEIN FOLDING;

D O I：

10.1007/BF01025477

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A plasmid analogous to the one described by Nagai and Thogersen (Nature, 309, 810-812, 1984) has been constructed for the expression of globins in E. coli. Induction with nalidixic acid produces high yields of a fusion protein, NS1-FX-beta-globin, where NS1 represents 81 residues of a flu virus protein and FX represents a blood-clotting Factor Xa recognition sequence, Ile-Glu-Gly-Arg. This fusion protein is readily solubilized in 50 mM NaOH and remains in solution when the pH is adjusted to 8.6. Under these conditions, the fusion protein is hydrolyzed by activated Factor X, giving authentic beta-globin which can be folded in the presence of cyanohemin and native alpha-chains to produce a tetrameric hemoglobin with the functional properties of natural human hemoglobin.

引用

页码：495 / 501

页数：7

共 21 条

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[2]

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