Bete-Galactosidase Protein Experiment

[JDAD]

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i am trying to find information on an interesting experiment to perform with the protein beta-galactosidase.

Does anyone have any ideas for me? I have already purified and isolated the protein, now I need an experiment that characterizes it.

Thanks

 

[Treg]

bGAL is often used as a reporter, so you can do a reaction where you add X-GAL + a few other things and turn cells blue. If you have purified it, you could do a colorimetric assay to determine its activity.

Hope that helps 🙂

Treg

 

[sweatybrain]

it's pretty well characterized. the only novel thing you can do with the protein (that I can think of off the top of my head given my general ignorance about this particular enzyme) is single molecule kinetics measurement....could be interesting to look at the difference between single molecule and "ensemble" kinetics.

Otherwise, just do some two hybrid in yeast to detect protein-protein interaction...but you don't need to purify the protein for that 🙂

 

[JDAD]

The fact that this protein is one of the most studied proteins ever makes it hard to do something unique.

I think I have decided on spectrophotometric determination of kinetic proterties, like Vo, Km, and Vmax.

The X-gal thing has already been done to death, and it doesn't tell me anything about the protein, other than is if function, it only tells me that it is present.

I think the kinetic stuff will be interesting, does anyone have a protocol for me?

 

[patzan]

nerds.

 

[VPDcurt]

why are you so obsessed with this one protein? i would try to find something else to study that people aren't heavily researching across the country. no one wants to read another B-Gal paper.

 

[Medikit]

The fact that this protein is one of the most studied proteins ever makes it hard to do something unique.

I think I have decided on spectrophotometric determination of kinetic proterties, like Vo, Km, and Vmax.

The X-gal thing has already been done to death, and it doesn't tell me anything about the protein, other than is if function, it only tells me that it is present.

I think the kinetic stuff will be interesting, does anyone have a protocol for me?

How about studying the kinetics as it relates to the substrate being used. Choose a couple of different galactosides and determine their kinetics and compare them to eachother. You could see what the effects are when using a bulky galactoside as the substrate compared to using something like lactose.