title: [High expression of rabbit sperm membrane protein rSP10 in Escherichia coli and preparation of its specific antisera].
abstract: To study the position and immunogenicity of rSP10, the rabbit rsp10 gene which did not include sequences coding for signal peptides was inserted into expression vector pET30a. An in-frame fusion protein was made such that a His6 stretch was produced at the N terminus of re-rSP10. High expression was obtained, the amount of re-rSP10 up to 67% in the total bacterial protein. The re-rSP10 was purified by DEAE chromatography and the yield of purified re-rSP10 was approximately 50 micrograms/mL of culture. Western blotting analysis of re-rSP10 with rabbit polyclonal sera raised against rabbit sperm membrane protein showed that the synthesized antigen possessed immunogenicity of rSP10. Specific antisera against re-rSP10 was induced using purified re-rSP10 as an antigen. The motility of capacitated sperms were affected but no aggregation was observed.
authors.affiliation: National Laboratories for Agrobiotechnology, China Agricultural University, Beijing 100094, China.
authors.forename: J X
authors.initials: JX
authors.lastname: Liu
authors.forename: A X
authors.initials: AX
authors.lastname: Lin
authors.forename: Y
authors.initials: Y
authors.lastname: Yang
authors.forename: J
authors.initials: J
authors.lastname: Hou
authors.forename: H Y
authors.initials: HY
authors.lastname: Hu
authors.forename: Y F
authors.initials: YF
authors.lastname: Chen
mesh_terms: D000818:Animals; D000954:Antigens, Surface; D004926:Escherichia coli / Q000235:genetics*; D005260:Female; D007106:Immune Sera / Q000276:immunology*; D008297:Male; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000276:immunology; D010957:Plasmids; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000276:immunology
chemical_list: D000954:Antigens, Surface; D007106:Immune Sera; D008565:Membrane Proteins; D011994:Recombinant Proteins; C049242:rSMP-B protein, human