Team:Botchan Lab Tokyo/BBa J04450
Botchan Lab. Tokyo
④RFP
please refer＜BBa_J04450＞
1. Long description
This parts is things that BBa_J04450 is inserted in psB1C3, and can express RFP because the promotor is also inserted in upstream of RFP. The introduced host Escherichia coli forms a colony in the selective medium and glows red.
2. Usage
The RFP gene part is mainly used as a reporter gene. In order to confirm whether this part works even under γ-ray irradiation, we introduced it into E. coli and cultured it under γ-ray irradiation to confirm its function.
3. characterization
We transformed the plasmid which RFP is inserted in psB1C3 into DH5α, and tested radiation tolerance. The following graph shows the result.Please refer to 2.Radiation irradiation experiment Protocol in the protocol for the procedure.
survival rate of E. coli (have plasmid RFP(BBa_J04450)inserted)
Figure1. The viability of E. coli DH5α against gamma irradiation
E. coli showed a significant decrease in viability between 50 Gy and 100 Gy versus 10 Gy. From this result, we concluded that E. coli is highly sensitive to gamma irradiation when it is irradiated with 50 and more doses of Gy (As for 0 Gy, an irradiated sample was as same as a non-irradiated group, so the survival rate was calculated as always 1).
Discussion
This graph shows that the number of cells is significantly reduced with respect to the intensity of radiation. That is, it was shown that E. coli (DH5α) having a plasmid inserted with RFP has no radiation resistance.
4.Refernce
Wan H, He J, Ju B, Yan T, Lam TJ, Gong Z., (2002)., Generation of two-color transgenic zebrafish using the green and red fluorescent protein reporter genes gfp and rfp., Mar Biotechnol (NY). 2002 Mar;4(2):146-54.