00:01
All right, so this question asks about the usage of gfp or green fluorescent protein, as well as specifically its limitations.
00:12
So what features of this protein gfp must be considered and tested before we try to use it in an experiment and try to interpret data coming from the experiment? and so starting from the top, so gfp, once again, stands for green fluorescent protein and its most common use is to tag or attach gfp to a protein of interest, also known as the target protein.
00:40
And what this allows us to do is to have that protein marked with essentially a bright green fluorescence marker, which we can detect after exposing the protein complex to ultraviolet light.
00:55
And so while this is very, very useful for imaging and for deterrent, concentrations or presence of protein and, you know, obtaining data regarding that using fluorescence, which is a relatively easy process.
01:12
It does have some limitations.
01:15
And one of the most prominent limitations that is most likely what the question is asking about is the fact that gfp chromophore formation, the chromophore is essentially the complex after it binds to the protein, occurs pretty slowly.
01:31
The duration of time for this green light to begin emitting from a target protein after being tagged is at least two hours.
01:42
So because of this time limitation as well as the fact that during this two hours or more, there are requirements of oxygen.
01:56
And this can pose some issues.
02:00
Some other possibilities for features of the protein that need to be considered and tested before interpreting any results would be the fact that if used in high temperatures, it requires oxygen...