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Method for linking a synthesized protein to its mRNA-DNA complex

S Liu1, G V Shivashankar, T Sano

  • 1NEC Research Institute, Princeton, NJ 08540, USA. shumo@research.nj.nec.com

Biotechniques
|November 1, 2000
PubMed

Insights

Preventing the termination of both DNA transcription and mRNA translation allows proteins to remain complexed with their mRNA and DNA. This finding supports in vitro protein synthesis and protein micro-array applications.

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Synthetic Biology

Background:

  • Nascent proteins can remain associated with ribosomes and mRNA if stop codons are deleted.
  • Messenger RNA (mRNA) forms stable complexes with DNA when transcription termination is blocked.

Purpose of the Study:

  • To investigate the principle that preventing both mRNA translation and DNA transcription termination can maintain a protein-mRNA-DNA complex.
  • To develop and test a method for creating such a complex.

Main Methods:

  • Designing a method to prevent both transcription termination and mRNA translation.
  • Utilizing an immobilized luciferase gene sequence for protein production.
  • Observing the association of the produced protein with its mRNA and DNA.

Main Results:

  • A functional luciferase protein was successfully produced.
  • The produced luciferase protein remained associated with its mRNA and DNA.
  • The feasibility of the proposed scheme for maintaining protein-mRNA-DNA complexes was confirmed.

Conclusions:

  • It is feasible to maintain a protein-mRNA-DNA complex by preventing the termination of both transcription and translation.
  • This method has potential applications in in vitro protein synthesis.
  • The technique could be applied to the development of protein micro-arrays.

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