Team:Tsinghua/Modeling

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Introduction

Phage production in E.coli

Naked Plasmid Vector Production

One of our team's sub project is the use nakes plasmid binded with a specified protein as the vector of gene therapy DNA. The normal replication process of plasmid will be changed due to the binding of specified protein(in our model,lambda repressor). Considering production of the plasmid protein complex, we want to maxize the amounts of plasmid protein complex, so a balance between the amounts of plasmid and protein is needed. In this model, we want to find out the optimized parameters for this plasmid vector production process.

Reaction kinetics

DNA protein complex’s binding and separating is a dynamical process

  • Eq 1.gif


The replication and degration of plasmid. The plasmid is considered to replicate in a constant rate (r) without the interference of binding protein. Here the parameter reff means the actual replicating rate of plasmid taken the binding protein into consideration.

  • Eq 3.gif


The transcription of λ repressor, notice that λ repressor is coded on the plasmid instead of in the genome of E.coli

  • Eq 4.gif

The translation of λ repressor

  • Eq 5.gif


Plasmid-protein complex affect plasmid replication

If a replication is already initiated but when the replication forks meet a lamda protein on the way, this replication will be aborted. So only when replication forks pass the protein binding point without lamda protein there, the replication processes are effective. This is a simplified model on how plasmid-protein complex affect plasmid replication, we will try to find out some more detailed mechanisms in the future. The probability of a plasmid to become a protein-DNA complex is:

  • Eqn6.gif

Because of the DNA-protein complex, the effective replication rate of plasmid is

  • Eqn7.gif

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