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).

<div class="ec--h2">Fusion protein for bone and cartilage regeneration</div>

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 (Zhou et al., 2016).

Zhou, N., Li, Q., Lin, X. et al. Cell Tissue Res (2016) 366: 101. https://doi.org/10.1007/s00441-016-2403-0

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_K2804004

CBD cipA fused to sfGFP

BBa_K2804005

CBD cipA fused to sfGFP under the control of LacI promoter

BBa_K2804014

CBD cipA fused to sfGFP

BBa_K2804015

CBD cipA fused to BMP2

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

<div class="ec--h2">Fusion protein to provide elasticity to the biomaterial</div>

Coding parts

BBa_K2804011

ELP V-150

BBa_K2804012

ELP V-150

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

<div class="ec--h2">Cellulose production module</div>

Coding parts

BBa_K2804008

CmcAx: Endo-beta-1,4-glucanase ezyme

BBa_K2804009

CcpAx: Cellulose-complementing protein

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

Team:Ecuador/Parts

PARTS

Index