Team:NCTU Formosa/Safe Project Design

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Safe Project Design

Choosing a non-pathogenic chassis

Bacterial strain: Escherichia coli K12

Escherichia coli K-12 are not a threat to human, animal (chickens, pigs, and calves), plants or the environment, according to the 1997 Final Risk Assessment of Environmental Protection Agency (EPA). “Any concerns in terms of health considerations are mitigated by its poor ability to disseminate, colonize the colon and establish infections in a murine model” (Smith et al., 2010). In addition, the chance of the insertion gene mutating the bacterial strain to be hazardous is minimal as the inserting genetic materials have to meet the EPA’s criteria (limited in size, well characterized, free of certain nucleotide sequences, and poor mobility).

Choosing parts that will not harm humans/ animals/ plants

Name	Natural Function	Acquired By	Notes
Cell lines
H292 (Mucoepidermoid Pulmonary Carcinoma cell)	Cell	Company
HCC827 (Adenocarcinoma)	Cell	Company
H1975 (Adenocarcinoma)	Cell	Company
MCF7 (Adenocarcinoma)	Cell	Company
MDA-MB-231(Adenocarcinoma)	Cell	Company
SK-BR-3 (Adenocarcinoma)	Cell	Company
Normal Backbone
pSB1A3	Plasmid backbone	Kit	http://parts.igem.org/Part:pSB1A3
pSB1K3	Plasmid backbone	Kit	http://parts.igem.org/Part:pSB1K3
pSB1C3	Plasmid backbone	Kit	http://parts.igem.org/Part:pSB1C3
Expression Backbone
pSB6A1	Plasmid backbone	Kit	http://parts.igem.org/Part:pSB6A1

Inserts:

Name	Natural Function	Acquired By	Notes
Promoters
J23101	Consitutive promoter	Kit	http://parts.igem.org/Part:BBa_J23101
J23110	Consitutive promoter	Kit	http://parts.igem.org/Part:BBa_J23110
R0010	Induced promoter	Kit	http://parts.igem.org/Part:BBa_R0010
Ribosome Binding Site (RBS)
B0034	Strong; efficiency 1.0	Kit	http://parts.igem.org/Part:BBa_B0034
B0030	Strong; efficiency 0.91	Kit	http://parts.igem.org/Part:BBa_B0030
Fluorescent Reporters
E0040	Green Fluorescent Protein	Kit	http://parts.igem.org/Part:BBa_E0040
E1010	Red Fluorescent Protein	Kit	http://parts.igem.org/Part:BBa_E1010
K592100	Blue Fluorescent Protein	Kit	http://parts.igem.org/Part:BBa_K592100
Chromoprotein
K592009	amilCP	Kit	http://parts.igem.org/Part:BBa_K592009
Transmembrane Protein
Lpp-OmpA fusion protein	Transmembrane protein in E.coli	Paper
FadL	Transmembrane protein in E.coli	Paper
Single-chain variable fragment (scFv) antibodies
scFv of Anti-EGFR	Part of Antibody	Drug bank
scFv of Anti-VEGF	Part of Antibody	Drug bank
scFv of Anti-HER2	Part of Antibody	Paper
Gold Binding Polypeptide (GBP)
Gold Binding Polypeptide		Paper
Terminator
J61048	Terminator	Kit	http://parts.igem.org/Part:BBa_J61048

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NCTU_Formosa APOllO
National Chaio Tung University
Engineering Building 6 EF455, 1001 University Road, Hsinchu 300, Taiwan, ROC.