全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...

Cloning, Soluble Expression and Characterization of Human sBCMA
人sBCMA cDNA的克隆、高可溶性融合表达及活性分析

Keywords: RT-PCR,sBCMA,fusion expression,proliferation inhibition
RT-PCR
,sBCMA,融合表达,增殖抑制

Full-Text   Cite this paper   Add to My Lib

Abstract:

BCMA is one of the transmembrane receptors belonging to BAFF and APRIL. In order to identify the feasibility of sBCMA as decoy receptor and obtain active sBCMA for its structural and functional research, full length of hBCMA was amplified with total RNA from Raji cell line by RT-PCR, and the cDNA encoding the extracelluar soluble domain of hBCMA was inserted into pET43.1a( ) vector. The recombinant vector pET43.1a( )-sBCMA was transformed into E.coli Origami B(DE3) pLyS which is helpful for disulfide bond construction of expression proteins. After IPTG induction, the recombinant protein was expressed as soluble fusion protein, sBCMA-NusA-His_6, and identified by western blotting. Then the target protein was purified by Ni -chelating Sepharose Fast Flow. The binding activity between recombinant sBCMA and BAFF was detected by ELISA. Also, Recombinant sBCMA inhibited proliferation of mouse B cell stimulating by rhsBAFF. It was proved that recombinant sBCMA has good bioactivity and the method to express those proteins rich in disulfide bond is feasible and effectual.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133