[Construction and expression of eukaryotic recombinant of mouse fertilin beta subunit in HEK293 cells].
OBJECTIVE
To construct the eukaryotic expression recombinant pSG. SS. C3d3. YL-Fbeta and analyze the expression of mouse fertilin beta subunit in HEK293 cells.


METHODS
The 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.


RESULTS
The recombinant vector pSG.SS.C3d3. YL-Fbeta could express Fbeta in HEK293 cells.


CONCLUSIONS
The expression of Fbeta in eukaryotic cells provides a foundation for further researches on the effect of high F, expression on fertilization process.
Sun
Zhong-yi
ZY
Department of Urology, Research Institute of Field Surgery, Daping Hospital, Third Military Medical University, Chongqing 400042, China. sun200801@hotmail.com
Jin
Feng-shuo
FS
Li
Yan-feng
YF
Zhang
Jun
J
D051722:ADAM Proteins / Q000096:biosynthesis* / Q000235:genetics; D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D005057:Eukaryotic Cells / Q000378:metabolism*; D000072196:Fertilins; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D008297:Male; D008562:Membrane Glycoproteins / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis; D014162:Transfection
D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; D051722:ADAM Proteins; C000606454:ADAM2 protein, human; C000606458:Adam2 protein, mouse; D000072196:Fertilins