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Related Concept Videos

Ribozymes02:47

Ribozymes

The term ribozyme is used for RNA that can act as an enzyme. Ribozymes are mainly found in selected viruses, bacteria, plant organelles, and lower eukaryotes. Ribozymes were first discovered in 1982 when Tom Cech’s laboratory observed Group I introns acting as enzymes. This was shortly followed by the discovery of another ribozyme, Ribonulcease P, by Sid Altman’s laboratory. Both Cech and Altman received the Nobel Prize in chemistry in 1989 for their work on ribozymes.
Ribozymes can be...
Ribozymes02:47

Ribozymes

The term ribozyme is used for RNA that can act as an enzyme. Ribozymes are mainly found in selected viruses, bacteria, plant organelles, and lower eukaryotes. Ribozymes were first discovered in 1982 when Tom Cech’s laboratory observed Group I introns acting as enzymes. This was shortly followed by the discovery of another ribozyme, Ribonulcease P, by Sid Altman’s laboratory. Both Cech and Altman received the Nobel Prize in chemistry in 1989 for their work on ribozymes.
Ribozymes can be...
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
Ribosome Profiling02:24

Ribosome Profiling

Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...
Tagging and Fusion Proteins01:24

Tagging and Fusion Proteins

Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...

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Related Experiment Video

Updated: Jun 27, 2026

Identification of Functional Protein Regions Through Chimeric Protein Construction
11:39

Identification of Functional Protein Regions Through Chimeric Protein Construction

Published on: January 8, 2019

Development of Chimeric Ribonuclease A Inhibitor for Molecular Biology Applications: SUMO Fusion as an Engineering

Dmitry Sukhov1,2,3, Tatyana Petrova1, Daria Kruglova1

  • 1DNA-Technology LLC, 117587 Moscow, Russia.

Current Issues in Molecular Biology
|June 26, 2026
PubMed
Summary

A novel chimeric RNase A inhibitor (SUMO-RI) offers enhanced storage stability for molecular biology applications. While effective, SUMO fusion slightly reduces thermal stability, presenting a practical handling advantage with a minor trade-off.

Keywords:
RNA stabilityRNase inhibitorRT-qPCRSUMO fusionchimeric proteinprotein engineeringprotein–protein interaction

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Last Updated: Jun 27, 2026

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Protein Engineering

Background:

  • Ribonuclease A (RNase A) poses a significant challenge in molecular biology techniques by degrading RNA.
  • Existing RNase inhibitors, like recombinant Rnh1, have limitations in storage stability.
  • Engineering RNase inhibitors is crucial for improving their practical utility in sensitive assays.

Purpose of the Study:

  • To engineer a chimeric RNase A inhibitor with enhanced stability.
  • To evaluate the performance of the engineered inhibitor in conditions mimicking real-time RT-PCR.
  • To assess the trade-offs associated with the engineering strategy.

Main Methods:

  • Construction of a chimeric protein by fusing a SUMO domain to the N-terminus of murine Rnh1 (creating SUMO-RI).
  • Functional assays to test RNA protection against RNase A degradation.
  • Comparison of SUMO-RI performance and stability against commercial RNase inhibitors and recombinant Rnh1.
  • Thermostability assays to determine the temperature limits of SUMO-RI.

Main Results:

  • SUMO-RI effectively inhibited RNase A, protecting RNA during simulated real-time RT-PCR.
  • Its performance was comparable to commercial RNase inhibitors.
  • SUMO-RI demonstrated significantly improved storage stability compared to recombinant Rnh1.
  • A reduction in thermostability was observed above approximately 47 °C due to SUMO fusion.

Conclusions:

  • SUMO fusion is a viable strategy for engineering RNase A inhibitors with improved practical handling and storage stability.
  • The chimeric SUMO-RI offers a balance between enhanced usability and a slight decrease in thermal resilience.
  • This engineered inhibitor provides a valuable tool for molecular biology applications requiring robust RNA protection.