[High expression of rabbit sperm membrane protein rSP10 in Escherichia coli and preparation of its specific antisera].
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.
Liu
J X
JX
National Laboratories for Agrobiotechnology, China Agricultural University, Beijing 100094, China.
Lin
A X
AX
Yang
Y
Y
Hou
J
J
Hu
H Y
HY
Chen
Y F
YF
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
D000954:Antigens, Surface; D007106:Immune Sera; D008565:Membrane Proteins; D011994:Recombinant Proteins; C049242:rSMP-B protein, human