Amino acid sequence

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kikkid
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Amino acid sequence

Post by kikkid » Tue Oct 02, 2007 1:57 am

Could someone please tell me what the easiest way to change an amino acid sequence is? (In an experimental situation)

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mith
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Post by mith » Tue Oct 02, 2007 2:05 am

Cut it up
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Post by kikkid » Tue Oct 02, 2007 2:08 am

Thank you! Once it is cut up (forgiveme for sounding ignorant) how is the new sequence formed? I mean, won't the amino acids just go right back to their original sequence unless something else is altered?

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mith
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Post by mith » Tue Oct 02, 2007 3:08 am

Well, think of it this way, if you drop a cup, it'll shatter into pieces; will the pieces ever come back together and form a cup?

You're adding water to break a peptide bond.


Another way to change the AA sequence is to change the DNA template from which it is made.
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Post by dna89 » Fri Oct 26, 2007 9:58 pm

yup, mith is right. site directed mutagenesis.

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Post by Jessy Springfield » Mon Oct 29, 2007 8:17 pm

mith wrote:Another way to change the AA sequence is to change the DNA template from which it is made.


So do you have any information on the processes in which this is performed?

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Post by mith » Mon Oct 29, 2007 8:32 pm

umm, DNA mutation?
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Post by Jessy Springfield » Mon Oct 29, 2007 9:20 pm

mith wrote:umm, DNA mutation?


yes, but by manipulation of specific regions?

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Post by mith » Mon Oct 29, 2007 9:22 pm

Well you could randomly blast UV rays at it till some sort of mutation occurs in that region, otherwise you could try using a virus to integrate itself into that region.
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Post by victor » Mon Oct 29, 2007 10:25 pm

Well, I think I've ever heard about the process in the seminar that I attended. The professor who presented his paper said that to make a peptide (of course not a protein, it's too long) with a desirable sequence from a protein, first we can hydrolize the peptide bond of the constituting protein into the corresponding amino-acids.
Next we can purify these amino-acids and bind them again one by one using the help of cellulose membrane and the Fmoc chemistry agent. The process is binding the amino-acid that you want into the cellulose membrane and then slowly add it one by one using an "activated amino-acid", an amino-acid which has been bound into the Fmoc chemistry agent. This Fmoc-bound amino acid will have an activated C-terminus, so then it will react with the amino group with the first amino-acid which is bound within the cellulose. This process is repeated again and again until we get the whole sequenced peptide as we want :D

Here is the schematic process:
Image
hope you like it :D
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mith
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Post by mith » Mon Oct 29, 2007 10:29 pm

a synthetic ribosome lol
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Post by victor » Mon Oct 29, 2007 10:35 pm

:lol: hope they can do that :lol:
Usually they (the professor and the team) only make a peptide which is consisted about 5 amino-acids, but the process is automated so they can get about 1 chip/day (1 chip is consisted about 8000 peptide molecules). They also found that these short peptides ara powerful agent to reduce cancer and they have tested to the leukemia and it works :D
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