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Enzyme-complemented Activatorsorbent Assay (ECASA): Genetic Engineering For Enzyme-linked Immunosorbent Assay-type Mercuric Ion Detection.
J. Klein, R. Mattes
Published 1998 · Chemistry, Medicine
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The sensor component of bacterial mercury resistance systems is the metalloregulatory protein MerR, which has nanomolar sensitivity and high selectivity for Hg(II). A fusion protein of MerR and the alpha-peptide part of beta-galactosidase (LacZalpha) was constructed by fusing the relevant genes. The protein exhibited both MerR functions and alpha-complementing activity to the inactive LacZDeltaM15 (M15) protein. The bifunctional character of the appropriate MerR-LacZalpha-complemented M15 protein (MerR-LacZalpha:M15 protein complex) was used to develop a Hg(II)-specific enzyme-complemented activatorsorbent assay. Hg(II) was immobilized and presented on a matrix taking advantage of the high affinity of Hg(II) to SH residues. The immobilized Hg(II) could be specifically detected down to the parts-per-billion level by quantifying the beta-galactosidase activity of the bound fusion protein complex.
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