Ivaylo

Psycho, thanks !!! I think that I got the idea.

Yeah, sure. NADH is consumed by Complex I but that's not what I am asking

Hello !! I am wondering about the electrochemical gradient and the proton pumps in mitochondria - Complexes I, III and IV, how is it created? Protons move from the matrix to the intermembrane space but how is this coupled with electron movement along the chain? Is it because the outer side of the membrane gets negatively charged and that's the driving force for proton movement or is it a confomational change in these complexes that displaces the protons... Many thanks !!!

Seems good to me, Thanks !!!

http://www.madsci.org/posts/archives/oct2001/1004108157.Mb.r.html Here I found in brief some useful information for the site specific mutagenesis. For more information, type "site specific mutagenesis" in google.

Ok, I will try to do it that way with cloning and selective mutations and I will try with UV light cause I am curious what's gonna happen. Thanks everybody for the discussion and special thanks to CharonY. I will try to find the book, it seems I'll need it.

Ecoli, Yes, I am working in the university.

If I induce a mutation in a gene, I can isolate the protein, but I want to see the mechanism this mutation will effect, so it should be in a living cell. I can use PCR and then expose to UV - that's how I can get a mutated gene, but how to place it in a living organism so I can see what is gonna happen? Mind my dumb questions, but I am learning now, I would really appreciate anw answers. P.S Is there any mechanism so I can mutate (let's say) a specific nucleotide in a gene - a selective mutation of some sort???

Hello!! I got a problem and the whole thing is a mess. I want to induce UV mutations in a gene and see how this protein is gonna work. How is this thing done?? How should I effect only the gene I am interested in??? Thanks !!!

I haven't done it, I am making a project and I am facing real difficulties combining the different techniques, that's my first project though and I am quite inexperienced. Concerning the mild detergent treatment, I think it works cause I found it on the internet, but donno what detergent is being used, for how long and so on, but I think it' s working. CharonY, please check my new topic, I am trying to induce UV mutations in a gene, but the whole thing is one big mess.

CharonY, thanks for your reply. I found that mild detergents are also being used to make channels in the membrane so the ABs can pass through it.

Hello !! I want to know the machanism of the protein labeling with antibodies. I want to label a protein that is being synthesized in the cell. I don't understand, how this thing is happening... How the antibodies pass the selective membrane?? Thank You.

Yes, in most bands you get a density gradient, but in some you don't. You see the tail that ends somewhere, then you see black space and then you see green strains of Dna again, like little green spots, but they are seen. Yes, it is for measuring DNA degradation. Yes, you cut the DNA with an agent and then you perform the electrophoresis and see the DNA "moving away" from the cell. http://cometassay.com/ here you can find more info. about it if you are interested

Hello again I was talking about the comets that you get when treating DNA with factors that cut the DNA molecule, you start the electrophoresis and then on fluorescent microscope you see the fragments that have been cut. http://library.wolfram.com/examples/cometassay/Images/index_gr_6.gif Hello, I am characterising comet tails. I am not using a specific software for that, just Photoshop. I got problems cause when calculating the lenght of the comet tail, there are some fragments that are far away from ... how to say it... the main tail, that is intensively green. What should I do? Should I include them in the lenght of the whole comet? I am doing this for the first time and I will apreciate help of any kind.

Hello, I am characterising comet tails. I am not using a specific software for that, just Photoshop. I got problems cause when calculating the lenght of the comet tail, there are some fragments that are far away from ... how to say it... the main tail, that is intensively green. What should I do? Should I include them in the lenght of the whole comet? I am doing this for the first time and I will apreciate help of any kind.

Hello! I want to know, when counting the charge let's say of a simple peptide 5 amino acids let's say, i count only the residues of the positive lysine- arginine and histidine and negative- aspartate glutamate? Thank you.

Yes, that is what I thought. I supposed that these alpha subunits bond to the 19S subunit or it must be something like that, but the main role is played by the 19S.

Well thanks anyway Bluenoise.

Bluenoise, i took the info. from wikipedia, just the link u gave but it is not clear... What are there regualtory particles?

Hello! I got a question. I can not understand what is the exact function of the alpha subunits in the proteasome. It is written that it maintains the structure of the proteasome and is regulated by "regulatory particles" what are these regulatory particles and isn;t the 19S subunit that is responsible for the recognition of the polyubiquitin and so on?Thank you.

Thank u very much !

Hello again. Well I got a problem... I am looking at the reaction which transforms succinyl CoA into succinate... catalyzed by succinate thiokinase. There is a molecule H20 that is not included in the scheme at the book. My professor said that there is a lot of water and hydrogen in the cell and that is why it is not included. If it is so, why do the cell need NAD.H as a special transporter of hydrogen. Well id u can enlighten the matter a little bit I would be greatful.

Thank you!

Hello, I got a problem. I don't know the difference between glycerol and glycerate, "ate" worries me. If someone can explain it i would be very grateful. Sorry bout the dumb question, i am learning the basics.