Purification of hybrid beta-galactosidase proteins encoded by phi X174 E phi lacZ and Escherichia coli prlA phi lacZ: a general method for the isolation of lacZ fusion polypeptides produced in low amounts.
A facile immunoaffinity chromatography method is described for the purification of lacZ fusion gene products. The method is general for any molecule antigenically related to beta-galactosidase and involves only a single step. We report its use to purify the products of lacZ fusions with a bacteriophage gene, phi X174E, and an Escherichia coli chromosomal gene, prlA. The hybrid protein products of both of these genes are membrane bound and present in very low molar amounts with respect to total cellular protein. Evidence is presented that substrate-affinity chromatography is not applicable to the isolation of low-level fusion proteins such as these.
Struck
D K
DK
Maratea
D
D
Young
R
R
D010584:Bacteriophage phi X 174 / Q000235:genetics; D002846:Chromatography, Affinity; D004274:DNA, Recombinant; D004926:Escherichia coli / Q000235:genetics; D005696:Galactosidases / Q000302:isolation & purification*; D005822:Genetic Vectors; D007163:Immunosorbent Techniques; D007763:Lac Operon; D001616:beta-Galactosidase / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*
D004274:DNA, Recombinant; D005696:Galactosidases; D001616:beta-Galactosidase