ABSTRACT
<p><b>OBJECTIVE</b>To construct the eukaryotic expression recombinant pSG. SS. C3d3. YL-Fbeta and analyze the expression of mouse fertilin beta subunit in HEK293 cells.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>The recombinant vector pSG.SS.C3d3. YL-Fbeta could express Fbeta in HEK293 cells.</p><p><b>CONCLUSION</b>The expression of Fbeta in eukaryotic cells provides a foundation for further researches on the effect of high F, expression on fertilization process.</p>
Subject(s)
Animals , Humans , Male , Mice , ADAM Proteins , Genetics , Blotting, Western , Cells, Cultured , Eukaryotic Cells , Metabolism , Fertilins , Gene Expression , Genetic Vectors , Membrane Glycoproteins , Genetics , Plasmids , Polymerase Chain Reaction , Recombinant Proteins , TransfectionABSTRACT
Fertilin beta plays an important role in fertilization by its disintegrin domain as a sperm-specific antigen. This paper reviews its structure, localization and roles in fertilization, and suggests that fertilin beta, as an important target antigen, has a very promising value in the development of human immunocontraceptive vaccine.