Expression of Pol-Gag in different viruses

  • #1

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I wanted to clear something up, if I wanted to express the Pol-Gag Proteins of HIV-1 in two different viruses would it be expressed by simply putting a promoter into front of the genes? Let's say I had a pseudo-typed HIV-1 Vector's genes and T4 structural genes would the T4 structure express pol-Gag by adding a promoter to the pol and gag genes, if they were all in the same plasmid?
 
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  • #2
Ygggdrasil
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I'm not sure I completely understand the question. HIV is a virus that infects humans and T4 is a phage that infects bacteria, so combining the two is somewhat unusual. If you had a plasmid that expressed the T4 structural genes and Gag-Pol on another promoter, you could certainly get the cell introduce the plasmid into to produce both sets of genes, but I don't think that you could get the T4 structural proteins to incorporate Gag-Pol into the T4 phages without first modifying the Gag-Pol gene in some way.
 
  • #3
I'm not sure I completely understand the question. HIV is a virus that infects humans and T4 is a phage that infects bacteria, so combining the two is somewhat unusual. If you had a plasmid that expressed the T4 structural genes and Gag-Pol on another promoter, you could certainly get the cell introduce the plasmid into to produce both sets of genes, but I don't think that you could get the T4 structural proteins to incorporate Gag-Pol into the T4 phages without first modifying the Gag-Pol gene in some way.
That's what I was wondering would the T4 Phages Incorporate Pol-Gag by making extra copies of pol-gag? Maybe I should ask this question what about a virus allows uptake proteins are there internal slots they fit into? I know it is a strange combination however it is necessary for a project of mine, that I am physically trying to do, I have T4 Bacteriophage cultures and the Plasmids for HIV-1, trying to make a device that uses both structures but it needs to not be lytic cycle the bacteriophages.

I have already expressed the Chickenpox caspid in a Pseudo-typed HIV-1 Vector however I don't want to try this until I know more about the specifications of Protein Uptake. I don't want to accidentally make a lytic Vector, this is dealing with a experiment of metamorphism in biological organisms.

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But you answered the original question that pol and gag would need to be modified to be used in a bacteriophage, so I thought I owed you a explaination, Thanks Ygggdrasil.
 
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  • #4
Ygggdrasil
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That's what I was wondering would the T4 Phages Incorporate Pol-Gag by making extra copies of pol-gag? Maybe I should ask this question what about a virus allows uptake proteins are there internal slots they fit into?
I think generally this would depend on the specifics of the virus you are manipulating. There has been a lot of work done on how to incorporate foreign proteins into bacteriophages from work on phage display techniques.

I have already expressed the Chickenpox caspid in a Pseudo-typed HIV-1 Vector however I don't want to try this until I know more about the specifications of Protein Uptake. I don't want to accidentally make a lytic Vector, this is dealing with a experiment of metamorphism in biological organisms.
Chickenpox, a virus that infects humans, is very different from T4, a phage that infects bacteria. It is unlikely that work on T4 would inform how to safely handle the viruses you create with the chickenpox capsid. Work using viral vectors, especially those that can infect humans, is hazardous, so universities generally require those handling viral vectors to get special training. Furthermore, such work generally needs to be performed in laboratory spaces certified to perform biosafely level-2 work. I would not recommend pseudotyping human viruses unless you are working in appropriate facilities and have the appropriate training.
 

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