replacing Cells in and outs

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mehdi71000
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Post by mehdi71000 » Sun Jun 03, 2007 11:47 am

Thank mate
I read the PDF and I found the products below in it. Witch one do you think is the glow vector?
Thanks

http://www.promega.com/catalog/catalogp ... say+System

mehdi71000
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Post by mehdi71000 » Sun Jun 03, 2007 12:17 pm

If I’m not mistaken
Luciferase Cell Culture Lysis 5X Reagent
Is suitable for plants and every thing else.

"The Luciferase Assay System may also be used for quantitation in plant and bacterial cells, but only Cell Culture Lysis Reagent is suitable for these applications"


Or is it best I should purchase Luciferase Assay System 100 assays. Witch I think has every cell in it. :?

I'll gives them a call tomorrow and keep you posted if you’re interested.
Thanks you all for your helpful replies

mehdi71000
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Post by mehdi71000 » Mon Jun 04, 2007 12:40 pm

ok this is the reply i ve got:
Thanks for contacting Promega technical services. I think that you may be a little confused about what you are after here. We have firefly luciferase vectors under the control of the strong viral SV40 promoter which would be able to express the luciferase gene in plants. However, they will not “glow” unless the substrate for luciferase is present as well. The substrate is luciferin and this is not expressed in plants so just expressing luciferase will not lead to any bioluminescence. The other thing to consider is that even if the luciferin is present the plants will not glow as the half life of the reaction is very short (2-5 minutes). Could I ask why you want such a vector and why you want to produce plants that will “glow in the dark”? I think you may be looking for a Green Fluorescent Protein (GFP) expressing vector. This vector have been used in the past to label cells and organisms and will produce a green fluorescence when excitation and emission are provided by the correct UV source.

:?

But I’m sure they have made glowing tobacco plants . Haven’t they?

david23
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Post by david23 » Mon Jun 04, 2007 1:03 pm

hahhaha, see how important it is to have GFP. Ask if they have cheap GFP. The only difference I see from the reply is that GFP glow lasts longer. Also ask if they have YFP and CFP as well. If you already bought the luciferase, keep it. You can use it for other stuff.

mehdi71000
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Post by mehdi71000 » Mon Jun 04, 2007 1:47 pm

What if I add luciferase expression vector + luciferin expression vector. So the luciferin is available for reaction. And since the cell keeps growing and making now enzymes and proteins the reaction will last? You think would it work?
They said the luciferase will work in mammalians but they said they'll get back to me on that.
I mean what’s the point of a fish that glows that it doesn’t glow in the dark. only under uv.
what can i use luciferase for?

david23
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Post by david23 » Mon Jun 04, 2007 2:01 pm

forget the UV thing. The half life thing is more of a concern but who cares about that. luciferin is added separately. No need for vector. Luciferin is not sold in vectors, it's not even a gene.

http://en.wikipedia.org/wiki/Luciferin

Look at the picture, it's not an amino acid, it's made separately by other mechanisms. To satisfy your glowing plants you would have to constantly add more and more luciferin each day. GFP dont have this problem. Save the luciferase for other experiments that you can play with later, if you want to do genetic engineering for real with those plants or your fish.

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Post by mehdi71000 » Mon Jun 04, 2007 2:13 pm

I see your point but there must be an enzyme or a protein that create this chemical. Any ideas what it might be?

david23
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Post by david23 » Mon Jun 04, 2007 2:21 pm

There are, but it's most likely a lot of enzymes and too many substrates involved. You are making this more difficult.

mehdi71000
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Post by mehdi71000 » Mon Jun 04, 2007 2:35 pm

No worries I’ll email some companies and ask them if they have such vector. it would be much easier to know the name of the enzymes responsible maybe they have a vector fully developed that caries several genes for all the enzymes witch produce luciferin, witch in results makes luciferin in cells

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Post by david23 » Mon Jun 04, 2007 2:39 pm

it's not that simple and not premade.

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Post by mehdi71000 » Mon Jun 04, 2007 5:56 pm

Ok I asked the company and they said that they don’t have the luciferin substrates expression vector. So I guess ill keep on looking

Ok this may sound silly and promise you don’t laugh, I don’t know much about DNA.
I read some ware last year so I’m not sure if I’m right. It said genes start 20 something letters before ATATA or something same. If this is true could one break the dna at points. In result every broken peace has 1 gene in it. and by adding all of these broken dna into a cell. What are the chances of the cell expressing the ones that it can? For instance. we are looking for a gene A but we have as a result of splitting the dna we have genes A B C D and F. so buy placing all of them into a Cell if the gene is for an enzyme witch can be expressed, that one will be expressed. And the some others will be expressed as well but the desired gene A did get expressed?so in result I may end up with a cell with the protein A but also with a muscle protean .I can feel I’m of course a but would it works?

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Post by mehdi71000 » Mon Jun 04, 2007 11:07 pm


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