how to make glowing bacteria

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mehdi71000
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how to make glowing bacteria

Post by mehdi71000 » Sat May 12, 2007 1:30 pm

Hi everybody.
I’m very interested in genetic engineering a bacteria to make it glow. Can you help me by giving me instruction to how to create one please?
And were to buy the materials please? I saw a kit on the internet that makes people able to do this at home for $99. But they can’t ship to UK because it has hazards material or some thing. So can you help me please? Thanks

david23
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Post by david23 » Sat May 12, 2007 4:16 pm

werent u going to buy some bacteria from that girl? Anyways, i dont know any kits, I just know the general experimental protocols and the equipment needed. Let me dig up the protocol for ya. It's several protocols, but since you are making this for fun, you can skip like a whole bunch of stuff on there.

You need some GFP or other glowing genes on a plasmid premade, and some competent bacteria cells, and you are almost there. Also make sure you have a micropippete, hmmm an incubator, if you can get one. some petri dishes to culture the bacteria, and of course the agar stuff for it to grow on, and some of those nutrience glucose supplement things. There are a couple more reagents out there, let me look for the list.

If you are in a hurry then search for heat shock protocols online.

mehdi71000
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Post by mehdi71000 » Mon May 14, 2007 12:29 am

Thanks
I searched on Google for heat shock and I find the procedure pretty interesting. I always thought virus’s was the tool scientist used to inject bacteria with the new vector. But I don’t really understand the procedure.
thanks for your time I really appreciate it.
Is there any enzymes I need to buy or I just need the GFP on a vector? cad can i use any bacteria? Could I use a plant seed? Or a fish egg? Or maybe my thumb? Only kidding.
thanks

david23
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Post by david23 » Mon May 14, 2007 7:53 pm

here is a very simple protocol on bacteria transformation http://faculty.plattsburgh.edu/donald.s ... ation.html

All you need is the plasmid/vector, no other enzymes this time. but make sure whatever medium you grow the bacteria corresponds to antibiotic resistant gene on your plasmid. Without that, your bacteria will not be forced to use the plasmid, and they will not take up the GFP gene.

Your first step is to select a plasmid.

mehdi71000
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Post by mehdi71000 » Tue May 15, 2007 1:27 am

Thanks. I looked up somewhere in the net and there were about $1500 for 1mg. is that the going rate?
One more thing is this method only valid for bacteria’s? Can I use a plant seed without the shell for instance, thanks?
What would happen if I inject this vector? Into a chicken egg and cover the hole. Would the chick glow? I read somewhere a chicken egg is a big cell is this true?
If this is true, would all the cells of the chick glow? I suppose the chick would be very hungry since GFP consumes lots of energy. I read somewhere for every photon of light GFP uses 3 energy rich molecules. But I could be wrong

david23
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Post by david23 » Wed May 16, 2007 2:37 am

hmmm i have never done it with animals or plants, but i have colleagues that I have. They said it was simple because they had a bunch undergraduates did in less than an hour with goldfish. I will ask them what they did. This time it's not with normal heat shock. It's more complicated. I think they just injected the DNA right into the fish embryo, probably did a lot. I dont think they had a lot of high tech equipment at the time. On tv you see people use the needle thing under the microscope, but the guy I know doesnt have any of that stuff. It worked.

Ok for plant seeds, it is hard. Not only the seed coat is hard to get into, but also plant cells have a tough cell wall. I am going to look around to see if there is some cheap method to do it.

The GFP vectors we got were cheaper, and we also borrowed a few times from the neighbors. Do you have any money to spare?

mehdi71000
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Post by mehdi71000 » Wed May 16, 2007 2:47 am

Yeh sure. Can you throw some compatible medium and some containers as well? by the way what other Florence proteins you have?

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