Construction and expression of eukaryotic recombinant of mouse fertilin beta subunit in HEK293 cells.
- Author:
Zhong-yi SUN
1
;
Feng-shuo JIN
;
Yan-feng LI
;
Jun ZHANG
Author Information
- Publication Type:Journal Article
- MeSH: ADAM Proteins; biosynthesis; genetics; Animals; Blotting, Western; Cells, Cultured; Eukaryotic Cells; metabolism; Fertilins; Gene Expression; Genetic Vectors; Humans; Male; Membrane Glycoproteins; biosynthesis; genetics; Mice; Plasmids; Polymerase Chain Reaction; Recombinant Proteins; biosynthesis; Transfection
- From: National Journal of Andrology 2007;13(4):297-301
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct the eukaryotic expression recombinant pSG. SS. C3d3. YL-Fbeta and analyze the expression of mouse fertilin beta subunit in HEK293 cells.
METHODSThe cDNA fragment expressing the disintegrin domain of mouse fertilin beta was obtained by PCR, and then inserted into the eukaryotic plasmid pSG. SS. C3d3. YL to get recombinant plasmid pSG. SS. C3d3. YL-Fbeta, which was transfected into the HEK293 cell line to express the target protein Fbeta after identified by restriction enzyme digestion. And then Fbeta was detected by indirect immunofluorescence through confocal laser scanning microscopy, Western blot, immunohistochemistry staining and flow cytometry assay.
RESULTSThe recombinant vector pSG.SS.C3d3. YL-Fbeta could express Fbeta in HEK293 cells.
CONCLUSIONThe expression of Fbeta in eukaryotic cells provides a foundation for further researches on the effect of high F, expression on fertilization process.
