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Related Concept Videos

Replication in Eukaryotes02:31

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In eukaryotic cells, DNA replication is highly conserved and tightly regulated. Multiple linear chromosomes must be duplicated with high fidelity before cell division, so there are many proteins that fulfill specialized roles in the replication process. Replication occurs in three phases: initiation, elongation, and termination, and ends with two complete sets of chromosomes in the nucleus.
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The eukaryotic promoter region is a segment of DNA located upstream of a gene. It contains an RNA polymerase binding site, a transcription start site, and several cis-regulatory sequences.  The proximal promoter region is located in the vicinity of the gene and has cis-regulatory sequences and the core promoter. The core promoter is the binding site for RNA polymerase and is usually located between -35 and +35 nucleotides from the transcription start site. The distal promoter regions are...
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RNA Polymerase (RNAP) is conserved in all animals, with bacterial, archaeal, and eukaryotic RNAPs sharing significant sequence, structural, and functional similarities. Among the three eukaryotic RNAPs, RNA Polymerase II is most similar to bacterial RNAP in terms of both structural organization and folding topologies of the enzyme subunits. However, these similarities are not reflected in their mechanism of action.
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The “tree of life” describes the evolution of life and the evolutionary relationships between organisms. The root of the tree is the common ancestor to all life on Earth. All other species radiate from this point, much like the branches of a tree. The numerous tips of these branches on the tree of life represent every living, or extant, species. Extinct species, which are species that no longer exist, can be found towards the center of the tree. Currently, these organisms, both...
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Certain biochemical processes, such as embryonic development and cell growth regulation, depend on the repression of specific genes. DNA binding proteins known as eukaryotic transcription inhibitors regulate the repression of gene expression in eukaryotes. The presence of these inhibitors at the required location and time in the cell is triggered by the presence of hormones and additional signals from other cells.
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Towards PacBio-based pan-eukaryote metabarcoding using full-length ITS sequences.

Leho Tedersoo1, Sten Anslan2

  • 1Institute of Ecology and Earth Sciences, University of Tartu, Estonia.

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Long-read sequencing improves species identification in microbial ecology. Optimized Pacific Biosciences protocols enhance resolution for eukaryotes, particularly Fungi and Stramenopila, overcoming short-read limitations.

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Area of Science:

  • Microbial Ecology
  • Molecular Biology
  • Genomics

Background:

  • High-throughput sequencing advances microbial ecology.
  • Short-read methods lack species-level resolution and identification certainty.
  • Accurate identification of eukaryotes, especially Fungi and Stramenopila, is crucial.

Purpose of the Study:

  • Optimize Pacific Biosciences long-read metabarcoding protocols.
  • Achieve species-level identification for all eukaryotes.
  • Focus on Fungi (Glomeromycota) and Stramenopila (Oomycota).

Main Methods:

  • Utilized Pacific Biosciences long-read sequencing.
  • Targeted the internal transcribed spacer (ITS region) and partial small subunit rRNA gene.
  • Tested protocols on composite soil samples and mock communities.

Main Results:

  • Proposed optimal degenerate primers: ITS9munngs + ITS4ngsUni for eukaryotes.
  • Demonstrated improved species-level resolution compared to short-read methods.
  • Evaluated the advantages and disadvantages of long-read eukaryotic identification.

Conclusions:

  • Optimized long-read protocols enable accurate species-level identification of eukaryotes.
  • This approach enhances the study of microbial communities, including Fungi and Stramenopila.
  • Long-read sequencing offers a promising solution for resolving eukaryotic diversity.