Cloning & Transforming Bacterial Vector for Protein Production

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Discussion Overview

The discussion revolves around the challenges faced in cloning and transforming a human protein-producing gene into a bacterial cell for commercial protein production. Participants explore potential reasons for the lack of protein synthesis despite following correct procedures, including the differences between bacterial and eukaryotic gene structures.

Discussion Character

  • Homework-related
  • Conceptual clarification
  • Debate/contested

Main Points Raised

  • One participant suggests that the plasmid may not be able to recombine with the particular gene, raising questions about compatibility.
  • Another participant points out the fundamental differences between bacterial and eukaryotic genes, implying that these differences could affect protein expression.
  • A later reply mentions that while it is possible to express human proteins in bacteria, certain modifications are necessary when starting from gene sequences, contrasting with the use of mRNA sequences.
  • Some participants reference the successful production of insulin in bacteria as an example, indicating that while challenges exist, successful outcomes are possible.

Areas of Agreement / Disagreement

Participants express differing views on the reasons for the lack of protein production, with some focusing on plasmid compatibility and others on the inherent differences between bacterial and eukaryotic genes. The discussion remains unresolved regarding the specific cause of the issue.

Contextual Notes

Limitations include the lack of detailed information on the specific gene being cloned, the type of plasmid used, and the conditions of the bacterial transformation process. There are also unresolved questions about the necessary modifications for successful protein expression.

Who May Find This Useful

This discussion may be useful for students and researchers interested in genetic engineering, molecular biology, and the practical applications of cloning techniques in protein production.

Raghav Gupta
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Homework Statement



Scientists isolated a clinically important protein producing gene from human cells and they want to clone it and transform into a bacterial cell, so that they can induce the protein synthesis and produce the protein commercially. But they are unable to get the protein even though all procedures are correct- they isolated the DNA then they amplified the desired gene and cloned into a bacterial vector and transformed into a bacterial cell. What may be the reason for protein is not getting produced in the bacterial cell.

Homework Equations


NA

The Attempt at a Solution


Is it that some plastmid is not able to recombine with particular gene and the same plasmid combine?
 
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Consider some of the differences between bacteria and eukaryotes. Do genes from one type of organism contain features that the other type of organism lacks?
 
Ygggdrasil said:
Consider some of the differences between bacteria and eukaryotes. Do genes from one type of organism contain features that the other type of organism lacks?
But I have read that scientists are able to produce insulin from bacteria by this cloning method.
There are many differences in bacteria and humans.
Human have a large size DNA and bacteria have small size.
 
Yes, it is possible to express human proteins in bacteria, but there are important changes that you must make if you are starting from the gene sequence (hint: you do not need to make these changes if you are starting from the mRNA sequence). Review the basic steps of transcription in bacteria and in eukaryotes, and you are likely to come across the answer.
 

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