Heterodimer protein of recombinant human bone morphogenetic protein and efficient expression and renaturation method of heterodimer protein

Abstract

The invention relates to a heterodimer protein of a recombinant human bone morphogenetic protein and an efficient expression and renaturation method of the heterodimer protein. The method comprises the following steps: linking a BMP (Bone Morphogenetic Protein) 7 gene with a BMP2 gene through DNA fragments encoding (GlyGlyGlyGlySer)3 by virtue of a fusion PCR technology, inserting fusion gene into a pHis-NusA plasmid so as to construct a recombinant plasmid, introducing the recombinant plasmid into escherichia coli BL21 (DE3), culturing at 37 DEG C, and inducing the expression by virtue of 0.5mM IPTG. The results show that the BMP7 / BMP2heterodimer protein can be efficiently expressed by virtue of the method, the expressed protein is present in a form of an inclusion body and a large number of renaturated proteins can be obtained by virtue of simple purification and renaturation of the inclusion body. According to the method, the operation for obtaining the BMP active protein is simple, the preparation cost is greatly reduced and the method is suitable for large-scale fermentation production and provides the possibility of clinical wide application of BMP in the treatment of bone damage.

Description

technical field [0001] The present invention relates to high-efficiency expression of BMP7 / BMP2 heterodimer protein by Escherichia coli prokaryotic expression system, and preparation of biologically active recombinant human bone morphogenetic protein BMP7 / BMP2 heterodimer protein through denaturation and dialysis renaturation new method. Background technique [0002] Bone morphogenetic protein (BMP) is a protein that was first discovered in 1965 and can induce normal and ectopic osteogenesis. Ability to induce new bone formation. At present, more than 20 BMPs have been identified and cloned, and they belong to the members of the transforming growth factor β (transforming growth factor, TGF-β) supergene family. BMP2 is one of the BMPs with the most extensive research and the strongest osteogenic activity . BMP7 is mainly expressed in bone and kidney. BMP7 has strong osteoinductive activity, can stimulate the proliferation of undifferentiated mesenchymal cells and different...

AI Technical Summary

Problems solved by technology

[0005] At present, BMP protein is generally obtained by extracting from bovine bone, which is costly, low in purity and easy to contaminate

And higher animals are immune to heterologous BMP, which will weaken the osteoinductive activity of BMP

The use of insect cells or animal cells takes a long time to culture, the culture conditions are harsh, and the cost is high, which is not conducive to large-scale production.

The price of BMP protein is too high, which increases the burden on patients and limits its wide application in clinic

Images