A rapid fluorescence bioassay for the determination of selenium on agar plates.
The essential trace element selenium (Se) is involved in the form of selenocysteine at the active site of several prokaryotic and eukaryotic proteins called selenoproteins. These proteins have recently attracted attention particularly in relation to their application to human health and new characteristics of the genetic code. We have recently described a selenium bioassay based on a recombinant DNA construct in which the expression of the lac' Z gene in Escherichia coli is proportionally and specifically driven by UGA-directed selenocysteine incorporation. Here we have further developed this bioassay for more rapid and sensitive detection and measurement of selenium that permits screening of the selenium status on agar plates. Again, the inclusion of selenium into the lac'Z-fusion product is reflected by the level of beta-galactosidase activity, which in turn is reflected by the intensity of fluorescence on agar plates. This fluorescing agent is a 4-methylumbelliferyl moiety which is released through the cleavage by the enzyme of 4-methylumbelliferyl-beta-D-galactoside. The intensity of the fluorescence is easily detected by uv irradiation and photographed by polaroid or video cameras.
Liu
Z
Z
Department of Molecular Biology, Hebrew University-Hadassah Medical School, Jerusalem, Israel.
Reches
M
M
Engelberg-Kulka
H
H
D000362:Agar; D001681:Biological Assay / Q000379:methods; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005456:Fluorescent Dyes; D005697:Galactosides / Q000737:chemistry; D006923:Hymecromone / Q000031:analogs & derivatives / Q000737:chemistry; D012643:Selenium / Q000032:analysis* / Q000097:blood; D012680:Sensitivity and Specificity; D001616:beta-Galactosidase / Q000737:chemistry
D005456:Fluorescent Dyes; D005697:Galactosides; D006923:Hymecromone; C026845:4-methylumbelliferyl-galactopyranoside; D000362:Agar; D001616:beta-Galactosidase; D012643:Selenium