How Do Plasmids Influence Bacterial Gene Expression and Antibiotic Resistance?

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Discussion Overview

The discussion focuses on the role of plasmids in bacterial gene expression and their influence on antibiotic resistance. Participants explore how plasmid DNA is expressed, its independence or dependence on chromosomal DNA, and the implications of antibiotic presence on plasmid copy number and function.

Discussion Character

  • Exploratory
  • Technical explanation
  • Conceptual clarification
  • Debate/contested

Main Points Raised

  • Some participants propose that plasmid gene expression is mostly independent but partly dependent on the bacterial host's expression apparatus, with plasmids having their own promoters and, in some cases, regulators.
  • Others argue that the effect of antibiotics on plasmid numbers depends on whether the plasmids carry antibiotic resistance genes and the type of origin of replication.
  • It is suggested that the presence of antibiotics may not affect plasmid numbers but could increase copy number if resistance genes are present.
  • Concerns are raised about the metabolic costs associated with having a large number of plasmids in a bacterial cell, which may vary based on plasmid size.
  • One participant questions whether plasmids have operon units similar to DNA, suggesting that cloning plasmids requires consideration of these units and their regulatory mechanisms to maximize plasmid copies during cloning.
  • Another participant notes that naturally occurring plasmids vary in size and gene content, with larger plasmids potentially containing multiple genes necessary for replication and partitioning.
  • Discussion includes the impact of modifications made to natural plasmids when developing cloning vectors, which may affect their functionality.
  • Copy number control of plasmids is mentioned, with examples of high and low copy number plasmids based on their origin of replication.

Areas of Agreement / Disagreement

Participants express multiple competing views regarding the independence of plasmid expression, the effects of antibiotics, and the implications of plasmid size and modification. The discussion remains unresolved on several points, particularly concerning the regulatory mechanisms of plasmids and their cloning applications.

Contextual Notes

Limitations include the need for further clarification on the specific mechanisms by which plasmids interact with chromosomal DNA and the conditions under which antibiotic resistance genes influence plasmid dynamics.

gemma786
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Hi
I want to know about the role which plasmids play in a baterial cell.How the genes which are coded in plasmid DNA expressed themselves ? Does the transcription of plasmid DNA takes place independently of chromosomal DNA I mean to say that do the plasmids have promotor sequence for RNA polymerase to bind or plasmid first bind to chromosomal DNA forming episome inorder to express the genes which it contanins ? AND how the presence of antibiotics in the environment influence the number of plamids in a bacterial cell ?
And what is the effect of having large number of plasmid of same type in a bacterial cell ?
Please help me to find answer to these questions.
Thanks.
 
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gemma786 said:
How the genes which are coded in plasmid DNA expressed themselves ?
Does the transcription of plasmid DNA takes place independently of chromosomal DNA I mean to say that do the plasmids have promotor sequence for RNA polymerase to bind or plasmid first bind to chromosomal DNA forming episome inorder to express the genes which it contanins ?

Basically, expression of plasmid gene is mostly independent but it is partly dependent on the bacterial host expression apparatus. The plasmid genes have their own promoter but, in most cases, they need the bacterial host RNA polymerase for their transcription. Some plasmids have their own regulators.

gemma786 said:
AND how the presence of antibiotics in the environment influence the number of plamids in a bacterial cell ?

It depends if the plasmids codes for an antibiotic resistance genes and if a large number of the gene product encoded by the resistance genes is needed for resistance to be achieved. It will also dependent on the type of origin of replication of the plasmid. The presence of antibiotics may have no effect on the number of plasmids but in some cases it may results in increase number of plasmid copy number if the plasmids has an antibiotic resistance gene.


gemma786 said:
And what is the effect of having large number of plasmid of same type in a bacterial cell ?

Again it depends if the plasmid is fairly large or small but a large number of plasmid will have a metabolic cost due to its large number of replication events.
 
Thanks. jansmith and sorry for replying late.
I was just trying to understand how plasmid can be used as a cloning vector.
And as you said plasmids containing a gene for antibiotic resistance can multiply their copies if surrounding medium contains antibiotic but you also said that plasmids can also have regulators now does this means to say that plasmids have operon units like dna ? It would mean that in order clonise a plasmid, we would have to take into consederation those operator units and the products which would lead to activation of those operon units , because we want to make as many copies of plasmid as we can in cloning process along with the desired gene that we have introduced in plasmid , isn't it ?
I am not sure about this . I need help !
 
In nature, plasmids that are very small will only have one gene, which might be required for their replication, but very large plasmid can contain several genes, including genes that are required for their replication and partitioning into the daughter cells.

For cloning vectors, most plasmids were derived from naturally occurring plasmids but the cloning vector went through major modification and removal of DNA with coding regions. In some case, these modification had major impacts.

Copy number of plasmids are often controlled through the origin of replication. For example, pUC19 or pUC18 are high copy numbers and their source plasmid is pMB1 is a low copy number plasmid. pUC19/18 have a point mutation in their origin of replication which allows for the high copy number but they also lack a rop gene.
 

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