Team:Chiba/Assay/store

Storage実験方法

Experiment:
Fig. 1に示すプラスミドでそれぞれ形質転換した大腸菌株BL21およびSHuffle® 株について，アラビノース存在化で培養しフェリチンの発現を誘導した後，クエン酸鉄/アスコルビン酸鉄存在下で培養し，培養後の菌密度（OD600）および，コロニー形成能力をそれぞれ測定した（Fig. 2 and 3）。

Detailed procedure

1.E. coli strain BL21 and SHuffle® were transformed with the plasmids shown in Fig. 1.
2.All transformants were inoculated into small (2 mL) culture and was shaken for 12h (at 37°C).
3.Inoculated into flesh media (2 mL), shaken for 3 hours.
4.We added L-arabinose into each sample (final conc. 0.2%). Then cultured for another 9 hours (at 37°C).
5.After 9 hours, we measured cell density (OD600) of every sample, and put 10^7 cell of E.coli to fresh medium containing various concentration of iron citrate or iron ascorbate.
6.Then we shaking cultured E.coli for 12 hours.
7.We got 1 mL aliquot from each sample and centrifuged with 7,000 rpm, at 25℃ for 2 minutes.
8.We wasted supernatant and resuspended with 1 mL saline solution (0.9% NaClaq), and we measured cell density (OD600).
9.5 μL of the diluted cell suspension was spotted on the LB agar medium and incubated for 12 hours. Approximately 107, 106, 105, 104, 103, and 102 cells were contained in the spots.

絵とかあるといいよね！！