Team:Nanjing China Bio/results

All parts that we make consist of an anaerobic promoter and a luciferase gene. The luciferase gene was cloned from the iGEM part K387009, which has a RBS region, luciferase gene and two terminals.
There are 17 anaerobic promoters cloned in two different ways. Firstly, we ask 8 anaerobic promoter from iGEM. Secondly, we cloned 9 anaerobic promoters from BL21 genomic DNA , one E.coli strain. The information of these promoters is from the paper Effect of anaerobic promoters on the microaerobic production of polyhydroxybutyrate (PHB) in recombinant Escherichia coli, and thanks to their great work.
The anaerobic promoters from iGEM parts are listed below

Parts	Name	Length	Plasmid	Resistance
BBa_K387003	PfdhF: fdhF promoter, a hypoxia inducible promoter	99bp	pSB1C3	C
BBa_K404106	phTERT promoter	459bp	pSB1C3	C/C
BBa_K227007	puc promoter	651 bp	pSB1K3	K
BBa_K376003	J6 Oxygen Sensitive Promoter	83 bp	pSB1C3	C
BBa_K239005	NarK promoter, Fnr activated under anaerobic conditions	139 bp	pUC57	A
BBa_K239006	Modified NarK promoter	89 bp	pUC57	A
BBa_K258005	Oxygen promoter-Vitreoscilla hemoglobin(VHb) promoter in E. coli.	137 bp	pSB1A2	A
BBa_J61043		pBca1020-r0040	pSB1A2	A

The promoters that are cloned from BL21 are linked with luciferase. The parts that we made are listed below.

parts	name	plasmid	resistance
BBa_K905000	PnirB	pSB1C3	C
BBa_K905001	Padhe	pSB1C3	C
BBa_K905002	PgadA	pSB1C3	C
BBa_K905003	PgadA	pSB1C3	C
BBa_K905004	PhdeA	pSB1C3	C
BBa_K905005	PhdeD	pSB1C3	C
BBa_K905006	PldhA	pSB1C3	C
BBa_K905007	PpflA	pSB1C3	C
BBa_K905008	PftnA	pSB1C3	C
BBa_K905009	ParcA	pSB1C3	C
BBa_K905010	PhTERT+RBS34+luciferase+dt	pSB1C3	C
BBa_K905011	Pnark+RBS34+luciferase+dt	pSB1C3	C

Then, we link hypoxia-induced nirB promoter to the plasmid pSB1C3, because we want more iGEMers can use the promoter to do their research.
Then we put plasmids that each has an anaerobic promoter and a luciferase gene into VNP20009 by electroporation. Then we tested the strength and anaerobic sensitivity of our promoters by measuring the fluorescence strength of each VNP strain that has a different promoter after the anaerobic culture.