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基于葡萄球菌蛋白A基因的基因融合载体。

Gene fusion vectors based on the gene for staphylococcal protein A.

作者信息

Uhlén M, Nilsson B, Guss B, Lindberg M, Gatenbeck S, Philipson L

出版信息

Gene. 1983 Sep;23(3):369-78. doi: 10.1016/0378-1119(83)90025-2.

Abstract

Two plasmid vectors, containing the gene coding for staphylococcal protein A and adapted for gene fusion, have been constructed. These vectors will allow fusion of any gene to the protein A gene, thus giving hybrid proteins which can be purified, in a one-step procedure, by IgG affinity chromatography. As an example of the practical use of such vectors, the protein A gene has been fused to the lacZ gene of Escherichia coli. E. coli strains containing such plasmids produce hybrid proteins with both IgG binding and beta-galactosidase activities. The hybrid protein(s) can be immobilized on IgG-Sepharose by its protein A moiety with high efficiency without losing its enzymatic activity and they can be eluted from the column by competitive elution with pure protein A. The fused protein(s) also binds to IgG-coated microtiter wells which means that the in vivo product can be used as an enzyme conjugate in ELISA tests.

摘要

构建了两种质粒载体,它们含有编码葡萄球菌蛋白A的基因并适用于基因融合。这些载体可使任何基因与蛋白A基因融合,从而产生杂合蛋白,这些杂合蛋白可通过IgG亲和层析一步法进行纯化。作为此类载体实际应用的一个例子,蛋白A基因已与大肠杆菌的lacZ基因融合。含有此类质粒的大肠杆菌菌株产生具有IgG结合活性和β-半乳糖苷酶活性的杂合蛋白。杂合蛋白可通过其蛋白A部分高效地固定在IgG-琼脂糖凝胶上,而不会丧失其酶活性,并且可以通过用纯蛋白A进行竞争性洗脱从柱上洗脱下来。融合蛋白还能与包被有IgG的微量滴定孔结合,这意味着体内产物可在ELISA试验中用作酶结合物。

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