Team:KCL UK/Parts

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    • Mussel Foot Proteins
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Parts Overview

Introduction:

Our project aim is to create a synthetic protein polymer to coat our spinal scaffold which will allow for strong adhesion of the PCL to the site of injury, as well as provide excellent mechanical strength to spine. Mfp-5 is one of the first proteins secreted by the mussel byssus and its natural role is to adhere to underwater substrates. We have taken the coding sequence of the beta(β) isoform of Pvfp-5 and created a genetic toolbox to enable the formation of a Pvfp-5β polymer in E.Coli. The 2020 KCL iGEM Team have designed a co-expression system to enable the in situ polymerisation of Pvfp-5β. Our parts collection consists of 2 BioBricks and 3 composites. All composites are the original work of KCL iGEM 2020 and we have used the tyrosinase sequence from the iGEM registry. Our two inserts are the Pvfp-5β and Tyrosinase coding sequences and these will form two separate vectors after being cloned into our plasmid backbone. Several design considerations have been made to ensure tight transcriptional control, genetic selectivity, high yield purification and minimal toxicity to the host cell. The two vectors will be expressed in parallel to form a polymer of Pvfp-5β in vivo.

Part Name	Part Number	Type	Description	Designer	Length
T7 Promoter	BBa_K3635004	Basic	T7 promoter Coding sequence	Emily Blundell	19 bp
Ribosome Binding Site	J61101	Basic	RBS Sequence	John Anderson	26 bp
Lac Operator	BBa_K3635003	Basic	Lac Operator Sequence	Emily Blundell	25 bp
Histidine Tag	BBa_K3635005	Basic	7xHis Tag	Emily Blundell	21 bp
pBluescript II SK+ f1 Ori	BBa_K3635006	Basic	Origin of Replication	Emily Blundell	306 bp
Ampicillin Resistance	BBa_K3635007	Basic	AmpR	Emily Blundell	966 bp
Pvfp-5β Coding Sequence	BBa_K3635000	Basic	Pvfp-5β Insert	Emily Blundell/Abigail Conner	528 bp
Tyrosinase Coding Sequence	BBa_K3033013	Coding	Tyrosinase Insert	Philip Naderev P.Lagniton	891 bp
Renervate Plasmid Backbone	BBa_K3635008	Composite	T7 promoter+RBS+lac operator+7xhis-tag+pBluescript II SK+ f1 ori+AmpR	Emily Blundell	2219 bp
Pvfp-5β Expression Vector	BBa_K3635009	Composite	T7 promoter+RBS+lac operator+Pvfp-5β+7xhis-tag+pBluescript II SK+ f1 ori+AmpR	Emily Blundell	2755 bp
Tyrosinase Expression Vector	BBa_K3635013	Composite	T7 promoter+RBS+lac operator+Tyrosinase+7xhis-tag+pBluescript II SK+ f1 ori+AmpR	Emily Blundell	3116 bp

Click below to see our parts:

1 / 4

Pvfp-5β DNA Insert

2 / 4

Tyrosinase DNA Insert

3 / 4

Renervate Plasmid Backbone

3 / 4

Pvfp-5β Expression Vector

4 / 4

Tyrosinase Expression Vector

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