Totipotent Stem Cell From Molecular Components - (Feb/01/2014 )
Q1 I am wanting to make an animal cell from DNA, RNA, Protein, and Fatty Acids as well as Glucose and other sugars. I am wondering. Should I start with the cell membrane or the nucleus and nucleolus(both are very important)?
Q2 The pace of DNA replication is 8 2/3 hours for every chromosome and 2097152 of each chromosome in 1 week. Is that just an estimate because I know that it is 8 2/3 hours per replication origin.
Q3 Once I know how many proteins of a particular type should I do this:
Go to BioGPS
Look up the proteins
Make sure I have human selected(since I am making a human stem cell)
Average the mRNA expression to get stem cell mRNA expression
Multiply by 100 to get the amount of protein
Q4 I have heard of amplicons used in PCR. Do I really need one to copy a whole gene or just primers, DNA, and enzymes?
So you are trying to be the first person to create life from a collection of components, and you are asking for advice here. Sounds like an exam question to me!
Yes I am trying to be the first person to make life from its components. Those 4 questions I have are relevant to this and not all are like an exam question(such as the one related to gene expression by strands of mRNA which is the easiest way)
And then to get the amount of ATP produced per day would I take that number of grams and divide it by grams/mole to get the number of moles and then multiply that by 6.022 * 10^23 for amount of glucose in terms of molecules and then multiply that by 36 ATP per glucose(40 are formed but 2 are used in glycolysis and 2 are used in NADH transport to mitochondria)?
You could for the glucose, but then it depends on the cell type as to how much glucose it would use per time. I also don't know how you would feed your potential single cell with the amount so as to keep the concentration constant (osmilarity effects)!
Note that both the glucose and ATP amounts are just estimates, they could be orders of magnitude out.
My cell culture medium has at first the vitamins and minerals, proteins, fatty acids, and sugars that every cell needs.
I could weigh out the glucose so that I get the exact number of grams and then put it in the culture medium. Also I could count how many cells are there or will be there given the length of the cell cycle so that I know what x is in x grams of glucose = amount of glucose per cell per day * n cells.
If my cell has a 24 hour cell cycle that would be nice since that means that I wouldn't have to feed the cells multiple times in 1 day.