Part:BBa_K2447500
Constituitive expression of Photosensitive EL222 bacterial protein
This part allows constitutive production of the light-sensitive bacterial photosensitive protein EL222, a blue light activated transcription factor. This part could prove useful in the design of optogenetic (blue-light) circuit to regulate protein expression.
BBa_K2447501 is a basic part, a sensor. It has been characterized using BBa_K2447501.
In this construct, the photosensitive bacterial protein is constructively produced. In the presence of blue light, EL222 is activated via dimerisation and is able to bind upstream of the pLuxI promoter, thereby allowing the recruitment of RNA polymerase to activate expression of RFP. In the absence of blue light, EL222 does not dimerise and is unable to bind DNA; hence showing minimal RFP expression.
Usage and Characterisation
This part is inserted into pBbE8k backbone and subsequently characterized in E. coli 10Beta and MG 1655. These cells are grown in LB medium (with kanamycin) at 37 degrees to reach OD600 of 0.1 before blue light is administered for 7 hours. For every hour, RFP readings were obtained and cross-compared with the control (no blue light).
Strong RFP expression was shown from both cell types (under blue-light condition) as compared to the control condition where no light was administered. By the 6th hour onwards, RFP productions had tripled for both cell types when compared to the control condition.
In short, this construct could prove useful for any group seeking to elucidate gene expression based on blue light as a physical stimulus.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 551
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 121
- 1000COMPATIBLE WITH RFC[1000]
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