[Soluble expression of recombinant human BMP6 in Escherichia coli and its purification and bioassay in vitro].
BMP6 is a potent protein for future treatment strategies of bone regeneration as it is a very important regulator of bone homeostasis. Active BMP6 is a dimer containing multidisulfide bonds and is a highly hydrophobic protein prone to aggregation. To obtain soluble and active BMP6 in Escherichia coli, we compared the effects of four N-terminal fusion tags (TRX, GST, MBP and CBD) and N-terminal His6-tag. The expression and solubility were tested under the different conditions (expression hosts, temperatures and inductor concentrations). A series of experiments leads to the finding that the placement of MBP before the BMP6 is best in availing the soluble expression of the protein. Our study alsodemonstrates that in E. coli BL21trxB(DE3) cytoplasm, which is a thioredoxin reductase mutant strain, soluble homodimeric BMP6 can be formed. The overexpressed MBP-BMP6 fusion protein is purified by chromatography, and shown to be functionally active.
Lei
Rongyue
R
Laboratory of Medical Molecular Genetics, College of Medicine, Nankai University, Tianjin 300071, China.
Qiao
Yuhuan
Y
Yan
Jidong
J
Yang
Shuang
S
Zhu
Tianhui
T
D055418:Bone Morphogenetic Protein 6 / Q000096:biosynthesis* / Q000235:genetics; D002352:Carrier Proteins / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D005822:Genetic Vectors; D006801:Humans; D058619:Maltose-Binding Proteins; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012995:Solubility; D014169:Transformation, Bacterial
C525816:BMP6 protein, human; D055418:Bone Morphogenetic Protein 6; D002352:Carrier Proteins; D058619:Maltose-Binding Proteins; D011993:Recombinant Fusion Proteins