Part:BBa_K1094400
Enhanced green fluorescent protein (eGFP)
A modified version of the green fluorescent protein (GFP) derived from Aequorea victoria with enhanced fluorescence properties. The sequence can be used as either reporter or fluorescent tag.
The part was cloned into the expression vector pJAM1786 via pDONR207 in E. coli, by using the Gateway system by Invitrogen. Colony PCR and gel electrophoresis was performed to ensure the insert was in the vector (shown in the seven leftmost lanes of fig. 1).
The restreaks of those seven colonies were inoculated into liquid culture and grown overnight alongside three E. coli cultures containing empty plasmid (pBBR1MCS-2). Fluorescence was measured in an opaque ELISA reader plate. The following dilutions were made: 1X, 2X, 5X, 10X and 50X. OD(600nm) was measured to ensure similar cell density. Excitation was carried out at 485 nm and emission was collected at 535 nm.
The mean values of the fluorescence can be found in the table below.
(The raw data can be found here)
All dilution showed significantly more fluorescence than the control cultures except for the 50X dilution. The mean of the difference to the control cultures is shown as a function of dilution below.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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