INTRODUCTION
The aim of our project is to obtain a novel bone and cartilage regeneration treatment. The path we take to carry out this was transforming E. coli to produce a fusion protein, conformed by a cellulose binding domain and a protein that induce chondrogenesis and osteogenesis, also a fusion protein conformed by CBD and an elastin like protein was design in order to obtain an elastic biomaterial. We also tried to produce cellulose in E. coli to have a better control in its production with a LacI repressed promoter. The parts are briefly described below (see the part page if more information is needed).
Fusion protein for bone and cartilage regeneration
Coding parts
BBa_K2804000
CBD cipA with a C-terminal linker
This part was design to try to improve the imperial college (2014) CBD cipA part BBa_K1321014, because the CBD N-terminal is where the protein binds to the cellulose and the linker could interfer with this interaction.
BBa_K2804006
Superfolder GFP with six-histidine tail
The adition of an histidine tail to part BBa_K1365020 was done because it allows the characterization of fusion proteins binding sites through atomic force microscopy, thanks to its affinity to the equipment nickel tip.
BBa_K2804002
Bone morphogenetic protein 2 (BMP2)
Bone morphogenetic protein 2 (BMP2), is a member of the transforming growth factor-β (TGF-β) super-family. It is involved in chondrogenesis and osteogenesis in vitro via the expression induction of Sox9, Runx2 and its downstream markers.1
Composite parts
BBa_K2804001
CBD cipA fused to BMP2 under the control of LacI promoter
Therapeutic protein coupled to the CBD whose expression is regulated by the presence of IPTG in the medium.
BBa_K2804005
CBD cipA fused to sfGFP under the control of LacI promoter
Name |
Description |
Type |
BBa_K2804000 |
CBD cipA with a C-terminal linker |
Coding |
BBa_K2804001 |
CBD cipA fused to BMP2 under the control of LacI promoter |
Composite |
BBa_K2804002 |
Bone morphogenetic protein 2 (BMP2) |
Coding |
BBa_K2804004 |
CBD cipA fused to sfGFP |
Composite |
BBa_K2804005 |
CBD cipA fused to sfGFP under the control of LacI promoter |
Composite |
BBa_K2804006 |
Superfolder GFP with six-histidine tail |
Coding |
BBa_K2804014 |
CBD cipA fused to sfGFP |
Composite |
BBa_K2804015 |
CBD cipA fused to BMP2 |
Composite |
Fusion protein to provide elasticity to the biomaterial
Coding parts
Composite part
BBa_K2804013
CBD cipA fused to ELP_C5 under the control of LacI promoter
Name |
Description |
Type |
BBa_K2804011 |
ELP V-150 |
Coding |
BBa_K2804012 |
ELP_C5 |
Coding |
BBa_K2804013 |
CBD cipA fused to ELP_C5 under the control of LacI promoter |
Composite |
Cellulose production module
Coding parts
Composite part
BBa_K2804010
Operon member of bacterial cellulose synthase complex: Cmcax and CcpAx
Name |
Description |
Type |
BBa_K2804008 |
CmcAx: Endo-beta-1,4-glucanase ezyme |
Coding |
BBa_K2804009 |
CcpAx: Cellulose-complementing protein |
Coding |
BBa_K2804010 |
Operon member of bacterial cellulose synthase complex: Cmcax and CcpAx |
Composite |
