Expression and purification of biologically active rat bone morphogenetic protein-4 produced as inclusion bodies in recombinant Escherichia coli

被引：15
|
作者：
Klösch, B
Fürst, W
Kneidinger, R
Schuller, M
Rupp, B
Banerjee, A
Redl, H
机构：
[1] Ludwig Boltzmann Inst Expt & Clin Traumatol, A-1200 Vienna, Austria
[2] Univ Med, Inst Pathol, A-8036 Graz, Austria
来源：
BIOTECHNOLOGY LETTERS | 2005年 / 27卷 / 20期
关键词：
bone morphogenetic protein-4; E; coli; purification; stabilization;
D O I：
10.1007/s10529-005-1794-x
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
Rat bone morphogenetic protein-4 (rBMP-4) cDNA was cloned from rat osteoblasts by RT-PCR and expressed in E. coli. Monomeric, dimeric and polymeric forms of recombinant rat BMP-4 (rrBMP-4) were obtained from inclusion bodies after solubilization with urea. The dimer was separated from the remaining polymer and host cell contaminants using size exclusion chromatography. Furthermore, purified rrBMP-4 was stabilized at low urea concentration (40 mM) and at pH 8.5 through the addition of bovine serum albumin. Both, rrBMP-4 dimer and polymer were biologically active as tested by the induction of alkaline phosphatase activity in MC3T3-E1 cells.
引用
收藏
页码：1559 / 1564
页数：6
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