Questions tagged [recombinant]

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7
votes
1answer
441 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
114 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
2k 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
46 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
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0answers
48 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
3answers
101 views

Vectors for genetic engineering

Well can a bacterium be used as a vector?I have learnt that Agrobacterium tumifacian can be used to deliver a gene of interest in plant cells but on a book in a multiple choice question it says that ...
2
votes
1answer
36 views

Pichia pastoris and ornithine

Can Pichia pastoris (species of yeast used for recombinant protein) produce the non-protein amino acid Ornithine? Or does it express ornithine at all?
2
votes
1answer
54 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
919 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
176 views

Why are two forward and reverse priming sites depicted here? what do they do?

This question is in regard to the baculovirus expression system. (source: amsbio.com) Why do we need to generate primers for the polyhedrin promoter and the baculovirus? Primers are needed to ...
1
vote
1answer
378 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
115 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
1answer
45 views

Recombination Data Set

So I was looking over some genetics question and came across this data set. In Fruit fly Drosophila melanogaster, there is a dominant gene b+ for grey body color and another dominant gene c+ for ...
1
vote
2answers
69 views

Help with gene linkage and mapping!

Here's an interesting question I've been trying to figure out: My annotations are in red. So far, I've figured out: We know 630 colonies survived when plated with ACD antibiotics...meaning they ...
1
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0answers
58 views

making antibodies rather than buying them

I need large amounts of anti-CD20 and anti-pan-cytokeratin antibodies. At DAKO, they go for >$1,500/mg, and I need >100mg, hence a purchase is out of question. Instead, I plan to make a homebrew ...
0
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1answer
123 views

Selection of recombinant host by color selection method

In recombinant DNA tech, To select a recombinant host from non-recombinants, we have different types of techniques available. The one I'm talking about is the color selection method with the use of ...
0
votes
1answer
63 views

Gene of interest number of copies in transformed K. pastoris

When transforming a K. pastoris strain, is there a way to predict how many copies of the gene will be integrated following the homologous recombination? More specifically is it something that a ...
0
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1answer
43 views

use of antibiotics in selection of good plasmids

I have not understood the role of antibiotic medium in filtering the bacteria containing the recombinant plasmids with the ones having normal plasmids. Since all the plasmids of a certain bacterial ...
0
votes
1answer
96 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 ...
0
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0answers
35 views

QA/monitoring in recombinant vaccine manufacturing

On a16z[0] podcast on vaccines[1], an interesting tidbit came up: vaccine manufacturing in cell cultures is expected to hit a major challenge in terms of quality control due to unpredictable behaviour ...
0
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0answers
14 views

effect of amino acid limitation in CHO cells

I would like to know the effect of individual amino acid and their role in CHO cell growth and their effects on their metabolic pathway if any. And also if there is an amino acid which has an effect ...
0
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0answers
104 views

Heteroduplex and Products of cleavage of Holliday Junction (Resolution) [duplicate]

In homologous recombination (Holliday model), the last process i.e resolution can occur in two ways (horizontl and vertical cuts) leading to formation of either "splice" or crossover products or "...