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Questions tagged [recombinant]

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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 ...
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1answer
167 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 ...
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Are there vectors available without any promoter?

Looking for a vector with the following characteristics: 1. Without any promoter 2. Selectable marker: Puromycin 3. Blunt end restriction sites 4. Transfection host: Human or Mouse cell lines.
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1answer
42 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 ...
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1answer
96 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 ...
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1answer
35 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?
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29 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 "...
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0answers
56 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 ...
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1answer
94 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 ...
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0answers
44 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 ...
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2answers
114 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 ...
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0answers
467 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 ...
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1answer
343 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 ...
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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 ...
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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 ...
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0answers
861 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?
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1answer
111 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 ...
2
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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 ...
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1answer
439 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 ...