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7
votes
1answer
339 views

Recombinant protein fraction in E. coli

If a protein is heterologously expressed in E. coli under the T7 promoter, what fraction of the total protein concentration in the cell is the heterologously expressed protein? What could be its ...
6
votes
1answer
100 views

Proteins that are strongly overproduced in E. coli and S. cerevisiae?

I'm looking for some pointers to proteins that produce at really gigantic levels in E. coli and yeast (S. cerevisiae). Can anyone point to some champion proteins? Even in inclusion bodies and non ...
4
votes
1answer
1k views

How to secrete a recombinant protein from E. coli?

What are some secretory pathways that can be used to secrete a recombinant bacterial enzyme from E. coli? I have a recombinant protein (29kDa) that I will express in E. coli BL21 cells. For now I ...
3
votes
1answer
38 views

Has any research lab done serious work to engineer new bacteria which assemble graphene wafers?

I was thinking about crazy uses for engineered bacteria. Nano-assembly of Graphene seems like a potentially excellent target for the technology. Have any research/full-blown labs worked on this? Any ...
3
votes
0answers
22 views

What are flagged primers? [closed]

I'm interested in amplifying a sequence for further use with Gibson Assembly. I want to create overhang regions in my DNA fragment so there would be complementarity to the plasmid I'm trying to insert ...
2
votes
1answer
49 views

What is the point of being selection-free?

I'm reading "Highly efficient endogenous human gene correction using designed zinc finger nucleases" by Urnov et al. They propose a way to use zinc finger proteins for gene therapy. They repeatedly ...
2
votes
0answers
283 views

Pink E.coli cell pellet. Reasons? [closed]

I've harvested IPTG induced E.coli BL21DE3 cells' suspension culture by spinning at 5000 rpm/15 min/4 degrees C. The pellet after spin looked pale pink colored. What could be the reasons?
1
vote
1answer
81 views

Recombination fraction( frequency) in Homozygotes

I have a question about the recombination frequency I hope you have time to answer. Background: If we look at homozygous individual A/A B/B which has been formed from the gametes AB and AB, this ...
1
vote
2answers
64 views

If a given DNA oligo is an aptamer, will the corresponding RNA oligo with the same sequence be an aptamer aswell?

Functionality of aptamers depends upon the oligo's sequence and secondary structure. So, if I take a DNA aptamer and make an RNA oligo of the same sequence (T replaced with U obviously), will this RNA ...
1
vote
0answers
47 views

Calculating the recombination factor of four point genetic cross?

So, I'm trying to work out the recombination factor (RF) from this set of data: Which shows the 16 phenotypes of progeny of a cross of Saccharomyces cerevisiae yeast. I know how to go about a three ...
0
votes
1answer
35 views

Relative concentration of enzyme vs reaction product

I was reading a paper in which a recombinant protein (His-6 tagged) is expressed in E Coli (BL21 DE3). The yield of the enzyme isolated from the culture is reported as 10-30 mg per L of bacterial ...