A bioassay based on recombinant DNA technology for determining selenium concentration.
The trace element selenium has recently attracted attention, particularly because (i) selenocysteine is involved in the active site of various prokaryotic and eukaryotic enzymes, some of which have a role in human health; (ii) selenocysteine incorporation into these proteins is coded by UGA codons; and (iii) as a result, selenocysteine is now considered to be the 21st amino acid in an expanded genetic code. Here, we built recombinant DNA constructs in which expression of the lac'Z gene is driven in Escherichia coli by UGA-directed selenocysteine incorporation. In this system, levels of beta-galactosidase activity are proportionally and specifically related to the presence and concentrations of several specific simple selenium derivatives. The system can thus be used as a sensitive bioassay for their determination. This bioassay is one of a few using recombinant DNA technology to provide a reporter for simple detection of a chemical trace element.
Reches
M
M
Department of Molecular Biology, Hebrew University-Hadassah Medical School, Jerusalem, Israel.
Zhao
C
C
Engelberg-Kulka
H
H
D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry; D001483:Base Sequence; D001681:Biological Assay / Q000379:methods*; D003062:Codon / Q000235:genetics; D004274:DNA, Recombinant; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005069:Evaluation Studies as Topic; D005560:Formate Dehydrogenases / Q000235:genetics; D005798:Genes, Bacterial; D017930:Genes, Reporter; D007763:Lac Operon; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012329:RNA, Bacterial / Q000737:chemistry / Q000235:genetics; D012643:Selenium / Q000032:analysis*; D017279:Selenocysteine / Q000378:metabolism
D001426:Bacterial Proteins; D003062:Codon; D004274:DNA, Recombinant; D012329:RNA, Bacterial; D017279:Selenocysteine; D005560:Formate Dehydrogenases; D012643:Selenium