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Eukayotic cell from scratch


caters

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If you manage to do that experimentally you are going to have your free pick of a tenured position in biology or biochemistry anywhere in the world....

 

Currently we are not able to re-create cell artificially from scratch. One of the major things we are able to do is e.g. exchanging the chromosomes (or complete nucleus in case of eukaryotes) but we still require a functional cell as basis.

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I have a culture medium with vitamins, minerals, and sugars. I have a question. I have my chromosomes from DNA extraction that I separated using several rounds of differential centrifugation. I have them in separate test tubes with DNA replication enzymes such as helicase and DNA polymerase as well as DNA ligase. I have put them in an incubator with human body settings. I know that after 1 day I will have this number of chromosomes:

2x1 + 2x2 + 2x3 + 2x4 + 2x5 + 2x6 + 2x7 + 2x8 + 2x9 + 2x10 + 2x11 + 2x12 + 2x13 + 2x14 + 2x15 + 2x16 + 2x17 + 2x18 + 2x19 + 2x20 + 2x21 + 2x22 + 2x23. How long does 1 round of DNA replication take. Will the total number of chromosomes be an even larger power of 2? How many rounds of DNA replication can occur within 24 hours. (this whole DNA replication thing is at average speed.


also will I have a different power of two of each chromosome if these chromosomes have these numbers of base pairs?:

Chromosome 1: 247,199,719

Chromosome 2: 242,751,149

Chromosome 3: 199,446,827

Chromosome 4: 191,263,063

Chromosome 5: 180,837,866

Chromosome 6: 170,896,993

Chromosome 7: 158,821,424

Chromosome 8: 146,274,826

Chromosome 9: 140,442,298

Chromosome 10: 135,374,737

Chromosome 11: 134,452,384

Chromosome 12: 132,289,534

Chromosome 13: 114,127,980

Chromosome 14: 106,360,585

Chromosome 15: 100,338,915

Chromosome 16: 88,822,254

Chromosome 17: 78,654,742

Chromosome 18: 76,117,153

Chromosome 19: 63,806,651

Chromosome 20: 62,435,965

Chromosome 21: 46,944,323

Chromosome 22: 49,528,953

X Chromosome: 154,913,754


Can somebody reply please?

Edited by caters
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  • 2 weeks later...

I have a culture medium with vitamins, minerals, and sugars. I have a question. I have my chromosomes from DNA extraction that I separated using several rounds of differential centrifugation. I have them in separate test tubes with DNA replication enzymes such as helicase and DNA polymerase as well as DNA ligase. I have put them in an incubator with human body settings. I know that after 1 day I will have this number of chromosomes:

2x1 + 2x2 + 2x3 + 2x4 + 2x5 + 2x6 + 2x7 + 2x8 + 2x9 + 2x10 + 2x11 + 2x12 + 2x13 + 2x14 + 2x15 + 2x16 + 2x17 + 2x18 + 2x19 + 2x20 + 2x21 + 2x22 + 2x23. How long does 1 round of DNA replication take. Will the total number of chromosomes be an even larger power of 2? How many rounds of DNA replication can occur within 24 hours. (this whole DNA replication thing is at average speed.

also will I have a different power of two of each chromosome if these chromosomes have these numbers of base pairs?:

Chromosome 1: 247,199,719

Chromosome 2: 242,751,149

Chromosome 3: 199,446,827

Chromosome 4: 191,263,063

Chromosome 5: 180,837,866

Chromosome 6: 170,896,993

Chromosome 7: 158,821,424

Chromosome 8: 146,274,826

Chromosome 9: 140,442,298

Chromosome 10: 135,374,737

Chromosome 11: 134,452,384

Chromosome 12: 132,289,534

Chromosome 13: 114,127,980

Chromosome 14: 106,360,585

Chromosome 15: 100,338,915

Chromosome 16: 88,822,254

Chromosome 17: 78,654,742

Chromosome 18: 76,117,153

Chromosome 19: 63,806,651

Chromosome 20: 62,435,965

Chromosome 21: 46,944,323

Chromosome 22: 49,528,953

X Chromosome: 154,913,754

Can somebody reply please?

Okay I now have the speeds of DNA polymerase 1 and DNA polymerase 3:

DNA polymerase 1: 10 bp/sec

DNA polymerase 3: 1000 bp/sec

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Time For Chromosome Replication

is that just an estimate because I know that it is 8 2/3 hours per replication origin. Now Proteinase K is used in DNA extraction to get rid of histones so I don’t have to worry about X inactivation till I start making histones. Hmm There are 10 base pairs per helical turn and there are 14 helical turns per nucleosome(DNA wrapped around histone) so that is 140 base pairs per nucleosome.

Blood vessel formation

Blood vessels are formed the same way all other parts of the body are. STEM CELLS and of course mitosis of existing cells.

Stem Cell

This cell I am working on is a totipotent stem cell so I can put in the proteins only expressed in the cells of blood vessels and it will arrange into a blood vessel.

Alternate Splicing

Those splicosomes are in the test tube with RNA polymerase 2 and ribosomes. Yes I am aware of alternate splicing. That is what makes different types of muscle well different. One codes for skeletal, the other smooth, the other cardiac. but smooth muscle also has genes that are not expressed in skeletal muscle that makes them respond to nerves involuntarily and cardiac muscle also has genes that are not expressed in smooth muscle or skeletal muscle.

Membrane Proteins

I am going to have a lot of proteins in the membrane just 30 so glycoproteins. The rest will be transport proteins. Speaking of transport proteins I am going to have H+ ion channels in the mitochondria to set that initial gradient.

Protein having to do with cancer

Yes TNF alpha is what causes programmed cell death of cancer, uncontrolled cell division(at least one of the factors in that), and differentiation into cancer cells.

Tissues having more than 1 cell type

Now Yes I know that most tissues are made up of more than 1 type of cell but for example the brain is all neurons. That is an organ and tissue with only 1 main type of cell but it has several subtypes.

Microarray

The microarray is so that I can compare the stem cell gene expression to other cell gene expressions.

Surgery Question

I will need every type of cell for that and I was thinking “Maybe Surgery on the macro scale(not too large of a sample from each organ) and then separation of types using a microscope and microtools will help me get ready for the microarray. The question is how do I know that I have gotten every type of cell in the sample from a particular organ and will the person die from the surgery?

Gene Expression

I know that people have different gene expression profiles even if they have the same genetic profiles so I would want to make sure that my nuclear DNA extraction, mitochondrial DNA extraction and surgery for the microarray are all from the same person.

My Plan to Start Life for the Stem Cell

You know how just millivolts of electricity can trigger things? Well that is how I plan to do that spark of life for the stem cell but first I will see if the ion channels are enough to trigger that and then apply an electrode similar to the one used in those neuron experiments to trigger it if neccasary.

Opinion about Nuclear Protein

All my nuclear proteins I do beleive will either stay in the nuclear pores or in the nucleoplasm(fluid similar to cytoplasm but only in the nucleus)

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Congratulations, you made all of the parts, now what? Life is much more complicated than the sum of its parts (DNA + proteins + membrane + golgi + etc. etc does not equate to a cell), cell physiology is much harder to understand and play around with than anatomy. Just because you put all the pieces a cell is made out of together doesn't mean you'll get a properly functioning cell. Cells respond to spacial and temporal cues with extremely complex signaling networks, how do you hope to control something like simple like energy metabolism just by putting the pieces of a cell together? You solved energy metabolism eh? How about the rest:

 

http://web.expasy.org/cgi-bin/pathways/show_thumbnails.pl

 

http://web.expasy.org/cgi-bin/pathways/show_thumbnails.pl?2

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I am going to apply millivolts of electricity in the same strength as a neuron needs for an action potential to make this cell work properly. Since it is a stem cell all it really does is normal cellular reactions and no extra.

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I think you're vastly underestimating all the things that go into living things, as well as misunderstanding how the development process works.

development at first is all stem cells and in some areas continues to be all stem cells such as the brain and heart and other organs that have smooth muscle as well as your skeletal muscle. In other areas though like the endothelium of arteries and veins it starts off as stem cells only and then stem cells still help but mitosis also occurs. I am not underestimating what goes in a cell. here is a list of importance of the major organelles:

1: Cell Membrane

2: Nucleus and Nucleolus

3: RER

4: golgi apparatus

5: SER

6: mitochondria

7: peroxisomes

8: Lysosomes

 

Here is a list of importance of the minor organelles

1: cytoskeleton

2: centrosome

that is the way I am planning to put in the organelles. should I do it in a different order.

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I think you're vastly underestimating all the things that go into living things, as well as misunderstanding how the development process works.

 

I think you underestimate the ability of people to underestimate complexity of biological processes.

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  • 2 weeks later...

Makes me laugh brother, go read and understand the complexity of the cellular physiology. Putting pile of parts does not make a working machine. Cell is even more complex than that, mentioned earlier cells needs specialized cues to function properly.

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