Difference between revisions of "Part:BBa K1993017"

Latest revision as of 07:55, 16 October 2016

eGFP

eGFP is a type of GFP derivatives by mutation. This mutation dramatically improved the spectral characteristics of GFP, resulting in increased fluorescence, photostability, and a shift of the major excitation peak to 488 nm, with the peak emission kept at 509 nm. This matched the spectral characteristics of commonly available FITC filter sets, increasing the practicality of use by the general researcher. [1] For example, eGFP allowed the practical use of GFPs in mammalian cells.

In our project, in order to ensure our engineered MSCs were locating in inflamed lesions, we introduced eGFP gene. Firstly, we acquired it from a synthetic constructed plasmid we purchased from Addgene(Figure 1). Secondly, we combined eGFP with internal ribosome entry site (IRES) and different chemokine receptors by Gateway technology and constructed various composite parts. Thirdly, we transduced each type of composite parts into MSCs. Finally, we confirmed whether eGFP were expressed in vitro (293FT cells and MSCs)(Figure 2 & Figure 3) and in vivo (animal models). For more details, click in BBa_K1993008 and BBa_K1993009

Figure 1 DNA of eGFP was shown in agarose gel electrophoresis.

Figure 2 eGFP was expressed in 293FT cells.

Figure 3 eGFP was expressed in MSCs.

References

[1] Pédelacq JD, Cabantous S, Tran T, Terwilliger TC, Waldo GS (Jan 2006). "Engineering and characterization of a superfolder green fluorescent protein". Nature Biotechnology. 24 (1): 79–88. Sequence and Features