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5 results for “inteins”
Massive intein content in Anaeramoeba reveals aspects of intein mobility in eukaryotes
<p>The spread of mobile genetic elements within and between the genomes of diverse organisms has greatly impacted biology across the tree of life. Inteins are an enigmatic type of mobile genetic element capable of self-splicing at the protein level. While inteins are generally rare in eukaryotes, we have discovered that three strains of the newly described eukaryotic microbe Anaeramoeba have many dozens of inteins encoded in their nuclear genomes. Genomic investigation shows that Anaeramoeba's inteins were acquired by lateral transfer from viruses and bacteria, and subsequently spread to many different genomic locations via allelic and, possibly, non-allelic homing. The promiscuous nature of inteins in Anaeramoeba makes them promising substrates for genetic engineering and biotechnological applications.</p>
Thermally controlled intein splicing of engineered DNA polymerases provides a robust and generalizable solution for accurate and sensitive molecular diagnostics
<p>DNA polymerases are essential for nucleic acid synthesis, cloning, sequencing and molecular diagnostics technologies. Conditional intein splicing is a powerful tool for controlling enzyme reactions. We have engineered a thermal switch into thermostable DNA polymerases from two structurally distinct polymerase families by inserting a thermally activated intein domain into a surface loop that is integral to the polymerase active site, thereby blocking DNA or RNA template access. The fusion proteins are inactive but retain their structures such that the intein excises during a heat pulse delivered at 70–80°C to generate spliced, active polymerases. This straightforward thermal activation step provides a highly effective, one-component 'hot-start' control of PCR reactions that enables accurate target amplification by minimizing unwanted by-products generated by off-target reactions. In one engineered enzyme, derived from <em>Thermus aquaticus</em> DNA polymerase, both DNA polymerase and reverse transcriptase activities are controlled by the intein, enabling single-reagent amplification of DNA and RNA under hot-start conditions. This engineered polymerase provides high-sensitivity detection for molecular diagnostics applications, amplifying 5–6 copies of the tested DNA and RNA targets with >95% certainty. The design principles used to engineer the inteins can be readily applied to construct other conditionally activated nucleic acid processing enzymes.</p>
Thermally controlled intein splicing of engineered DNA polymerases provides a robust and generalizable solution for accurate and sensitive molecular diagnostics
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Massive intein content in Anaeramoeba reveals aspects of intein mobility in eukaryotes
Open the record for dataset details and reuse information.
MD trajectories for intein simulations
<p>MD simulation trajectories and run files for 400ns simulations for MchDnaB1-HAA. MchDnaB1-HN, and gp41-1WCT-HN inteins. The simulations were run with and without the N-extein sequence.</p>
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