[Cloning and expression of soluble B lymphocyte stimulator].
OBJECTIVE
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.


CONCLUSIONS
sBLyS with biological activity was successfully expressed and purified.
Sun
Jian
J
Department of Molecular and Cellular Pharmacology, Tianjin University, Tianjin 300072, China. jsun@public3.bta.net.cn
Li
Yan
Y
Feng
Jian-nan
JN
Sun
Ying-xun
YX
Hu
Mei-ru
MR
Shen
Bei-feng
BF
D000818:Animals; D053264:B-Cell Activating Factor / Q000235:genetics* / Q000378:metabolism*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000032:analysis; D005784:Gene Amplification; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000737:chemistry* / Q000378:metabolism; D016133:Polymerase Chain Reaction
D053264:B-Cell Activating Factor; D018076:DNA, Complementary