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Revision as of 00:43, 18 October 2018

iGEM ECUADOR
C-lastin, Interlab

PARTS

Index

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 (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

 

Fusion protein to provide elasticity to the biomaterial
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

 

Cellulose production module
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
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