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[Cloning and expression of soluble B lymphocyte stimulator]
Jian Sun1, Yan Li, Jian-nan Feng
1Department of Molecular and Cellular Pharmacology, Tianjin University, Tianjin 300072, China. jsun@public3.bta.net.cn
Aim:
To clone and express soluble B lymphocyte stimulator (sBLyS).
Methods:
Total RNA was isolated from peripheral blood mononuclear cells, and used to synthesize cDNA by reverse transcription. sBLyS cDNA was amplified by PCR with specific primers and inserted into a prokaryotic expression vector pET-30a. Recombinant plasmid was transformed into E.coli strain BL21(DE3). sBLyS was expressed in E.coli, purified in vitro, and analyzed with peptide mass fingerprinting and Daudi cell proliferation assay.
Results:
sBLyS cDNA was cloned. Peptide mass fingerprinting of purified BLyS matched with that of BLyS proteins. Purified sBLyS could stimulate Daudi cell proliferation in vitro.
Conclusion:
sBLyS with biological activity was successfully expressed and purified.
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