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plasmids

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plasmids

Postby mehdi71000 » Wed Jun 27, 2007 11:37 pm

Hi
From what I understand you can insert any plasmid on to any cell and it will get expressed but not always regulated. Is this correct :?
Does regulation means DNA is duplicated during cell duplication?
Thanks
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Postby canalon » Thu Jun 28, 2007 2:50 am

No. Regulation is at the gene level. It is about a gene will be expressed (ie, how often mRNA will be transcribed). But before even that happen you need your plasmid to be replicated in the cell and with the cell so that when your cells are replicating, the plasmid do not get lost.

Of course many wild types bacteria (and other cells too) use methylation of DNA to recognize exogenous DNA and destroy it.

So definitely not that easy to transform any plasmid into any bacteria.
Patrick

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Postby mehdi71000 » Thu Jun 28, 2007 9:43 am

Cheers mate for your knowledgeable wisdom

What is responsible for Dna duplication?

How can I tell witch plasmid can be expressed in witch organism? for example the one below
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Postby khenwood » Thu Jun 28, 2007 6:41 pm

Mehdi - You were asking similar questions on another forum. Actually I believe you asked the exact same question regarding this plasmid which raised concern from many forum members.

Can I ask why you're trying to do a genetically-engineered animal experiment yourself, if in fact you are asking basic genetic-engineering questions in an online forum?

My tone isn't meant to be offensive by the way.
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Postby siroma » Thu Jun 28, 2007 10:14 pm

mehdi71000 wrote:What is responsible for Dna duplication?

How can I tell witch plasmid can be expressed in witch organism? for example the one below


Ori of the plasmid is the only responsible element for duplication. One has Oris of virii, bacteria and yeasts (YAC).
Copy number maybe regulated in numerous ways.
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Postby mehdi71000 » Thu Jun 28, 2007 10:21 pm

No problem mate
I bought a glow in the dark transformation kit from Carolina science supplies I tried doing an experiment on e coli and it worked. I’m interested in genetic engineering. And I like to learn the basics. I wanted to try this experiment on plants but I realised its not easy as it seams. I require some sort of virus for the injection and that’s very difficult too perform .I mean glow fish has been done before. I love this subject and I can’t wait to start university next year hopefully. I love experiments and I love animals.
I hope that cleared you thoughts some how.
Now a question for you
Are you cop? If you are you have to tell me if not its entrapment
:D you sound like FBI :wink:
Last edited by mehdi71000 on Thu Jun 28, 2007 10:26 pm, edited 1 time in total.
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Postby mehdi71000 » Thu Jun 28, 2007 10:25 pm

Thanks siroma
Can the direction of the plasmid show witch spices the plasmid can be used on?
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Postby canalon » Fri Jun 29, 2007 3:11 am

mehdi71000 wrote:Thanks siroma
Can the direction of the plasmid show witch spices the plasmid can be used on?


Thyme obviously! It takes a lot of thyme to do good transformation ;)

There is no direction for the plasmid. But regulators have to be located upstream of the genes, so they have a "direction" if you want to insert the gene to express at the rigt spot.
Now if you look at your map you will see a small band labeled "ori", that is the origin of replication that will define which species can propagate the plasmid. Usually when nothing is written, it is either because the host is clear enough (you are looking at the insect vector page of a molecular biology catalog) or because it is E. coli and probably many other Enterobacteria.
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Postby mehdi71000 » Fri Jun 29, 2007 12:16 pm

Thanks mate you’re a genius.
Can you show me what are the other origins are called?
Just one more thing, I’ve heard of universal expression vectors are they the shuttle vectors you were talking about?
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Postby khenwood » Fri Jun 29, 2007 2:05 pm

Hahaha, don't let the "Washington DC" part throw you.

I'm not a cop or FBI, just a curious person. :)
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Postby mehdi71000 » Fri Jun 29, 2007 2:56 pm

You got it 1first time
Can you design me this plasmid?
I’ve tried registering on your forum but no reply yet. Can you ask on your forum can some one design me this plasmid. I’ll pay them good money
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Postby khenwood » Fri Jun 29, 2007 5:23 pm

No, I can't design or manufacture the plasmid for you.

In regards to the organization I work for, you have to be a qualified life science professional who is currently working. Once approved, members post in our forums.
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