Determining the color of the fluorescent protein
encoded in provided plasmid by implementing it into
E. coli BL21DE3.
, Abstract:
Fluorescent proteins are proteins that can emit fluorescence under ultraviolet light. In this
report, it will be determined which fluorescent protein was encoded in a bacterial colony of
Escherichia coli referred to as plasmid 3. To determine this, the plasmid is investigated at a
genome level and later on the protein level. At the genome level, the DNA was analyzed with
an agarose gel, the five visible bands found are similar to the banding pattern of Yellow
Fluorescence Protein (YFP). At the protein level, bacteria were grown, harvested and lysed
to analyze the protein. Afterwards, the molecular weight of the fluorescent protein was
determined by adsorption and fluorescent spectra. A molecular weight of 27,1 kDa was
found which lies close to the molecular weight of the YFP of 27 kDa. The measured
excitation and emission peaks during the spectrophotometry were 515 nm and 529 nm,
which are similar values to the 513 nm excitation peak and the 527 nm emission peak of
YFP. All these results led to the conclusion that the YFP was encoded into the DNA of
Plasmid 3.
encoded in provided plasmid by implementing it into
E. coli BL21DE3.
, Abstract:
Fluorescent proteins are proteins that can emit fluorescence under ultraviolet light. In this
report, it will be determined which fluorescent protein was encoded in a bacterial colony of
Escherichia coli referred to as plasmid 3. To determine this, the plasmid is investigated at a
genome level and later on the protein level. At the genome level, the DNA was analyzed with
an agarose gel, the five visible bands found are similar to the banding pattern of Yellow
Fluorescence Protein (YFP). At the protein level, bacteria were grown, harvested and lysed
to analyze the protein. Afterwards, the molecular weight of the fluorescent protein was
determined by adsorption and fluorescent spectra. A molecular weight of 27,1 kDa was
found which lies close to the molecular weight of the YFP of 27 kDa. The measured
excitation and emission peaks during the spectrophotometry were 515 nm and 529 nm,
which are similar values to the 513 nm excitation peak and the 527 nm emission peak of
YFP. All these results led to the conclusion that the YFP was encoded into the DNA of
Plasmid 3.
