ABSTRACT
To construct the eukaryotic expression plasmid of human PRX3 and measure its expression in the HEK-293FT cells, the full-length coding region of human PRX3 was cloned by PCR and inserted into the eukaryotic expression vector pcDNA4-Xpress (A). HEK-293FT cells were transiently transfected with the recombinant plasmid. Western blot and immuofluorescence were used to detect the expression of the fusion protein. In the experiment, restriction analysis identified the construction of the recombinant plasmid and the inserted sequence was identical with that published on GenBank. Western blot and immunofluorescence confirmed the expression of the recombinant protein in transfected HEK-293FT cells. It was concluded that the eukaryotic expression plasmid of human PRX3 was constructed successfully and the recombinant could be expressed efficiently in HEK-293FT cells, which provides a sound basis for the further study on human PRX3.
Subject(s)
Cell Line, Transformed , Cloning, Molecular , Embryonic Structures , Eukaryotic Cells/metabolism , Gene Expression , Genetic Vectors , Kidney/cytology , Kidney/metabolism , Peroxidases/biosynthesis , Peroxidases/genetics , Plasmids/genetics , TransfectionABSTRACT
Salivary lactate dehydrogenase (LDH), peroxidase (Px) and leucine aminopeptidase (LAP) activities were scanned in both normally ovulating and anovulating women during entire menstrual cycle. In ovulating women, all the three enzymes exhibited significant increase in the activity on or before the onset of ovulation which was monitored by the shift of the basal body temperature (BBT) as well as the ferning pattern of the cervical mucus. The peak maximum at the midcycle was several times higher than the previous day value in all the six normal women. In anovulatory women, no such remarkable change in the enzyme activities was found throughout the cycle. Salivary LDH and LAP showed peak at the midcycle and at the same time required short time for assay, so the present results are strongly suggestive that the determination of salivary enzyme content may be a convenient method for detecting the day of ovulation.