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Related Concept Videos

LTR Retrotransposons03:08

LTR Retrotransposons

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LTR retrotransposons are class I transposable elements with long terminal repeats flanking an internal coding region. These elements are less abundant in mammals compared to other class I transposable elements. About 8 percent of human genomic DNA comprises LTR retrotransposons. Some of the common examples of LTR retrotransposons are Ty elements in yeast and Copia elements in Drosophila.
The internal coding region of LTR retrotransposons and their mechanism of transposition closely resembles a...
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Non-LTR Retrotransposons03:18

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As the name suggests, non-LTR retrotransposons lack the long terminal repeats characteristic of the LTR retrotransposons. Additionally, both LTR and non-LTR retrotransposons use distinct mechanisms of mobilization. Non-LTR retrotransposons are further divided into two classes - Long interspersed nuclear elements (LINEs) and short interspersed nuclear elements (SINEs), both of which occur abundantly in most mammals, including humans. Some of the active non-LTR retrotransposons in humans are L1...
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Transposons make up a significant part of genomes of various organisms. Therefore, it is believed that transposition played a major evolutionary role in speciation by changing genome sizes and modifying gene expression patterns. For example, in bacteria, transposition can lead to conferring antibiotic resistance. Movement of transposable elements within the genetic pool of pathogenic bacteria can aid in transfer of antibiotic-resistant genetic elements. In eukaryotes, transposons can carry out...
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DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
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Translesion DNA Polymerases02:10

Translesion DNA Polymerases

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Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
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Bidirectional Retroviral Integration Site PCR Methodology and Quantitative Data Analysis Workflow
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Ty retrotransposon element based multiple integration toolkit for Saccharomyces cerevisiae.

Song Gao1,2,3,4, Weizhu Zeng1,3,4, Dong Li1,3,4

  • 1Engineering Research Center of Ministry of Education on Food Synthetic Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu, 214122, China.

Synthetic and Systems Biotechnology
|May 19, 2025
PubMed
Summary

This study introduces a novel toolkit for high-level protein production in Saccharomyces cerevisiae (S. cerevisiae) using Ty element integration. This method enables stable, multi-copy gene expression for enhanced protein and metabolite yields.

Keywords:
Gene overexpressionHigh copy number integrationMetabolic engineeringProtein productionRetrotransposons

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Area of Science:

  • Synthetic Biology
  • Molecular Biology
  • Biotechnology

Background:

  • Achieving high-level protein and metabolite production in Saccharomyces cerevisiae (S. cerevisiae) requires stable, efficient gene expression.
  • Genome integration of specific genes offers a robust strategy for sustained, high-level protein production.

Purpose of the Study:

  • To develop and validate a toolkit for extra-high-level gene expression in S. cerevisiae by integrating genes into Ty elements.
  • To demonstrate the efficacy of this toolkit for producing fluorescent proteins and a specific metabolite (taxifolin).

Main Methods:

  • Utilized five families of Ty elements in S. cerevisiae CEN.PK2-1D for massive gene expression.
  • Engineered nine selective markers, with six (TRP1, LEU2, URA3, HIS5, natMX, hphMX) enabling stable high-copy integration (>15 copies) at Ty sites.
  • Verified the toolkit by overexpressing fluorescence proteins and taxifolin biosynthesis pathway genes.

Main Results:

  • Achieved a protein titer of 1.6 g/L (268.1 mg/g DCW) for phiYFP, with 3.3 times higher fluorescence intensity compared to episomal overexpression.
  • Successfully integrated 14 genes for taxifolin biosynthesis into three Ty sites using three selective markers.
  • Accumulated 277.6 mg/L of taxifolin from glucose.

Conclusions:

  • The developed toolkit enables stable, high-copy integration of genes into Ty sites for enhanced protein and metabolite production in S. cerevisiae.
  • This approach significantly improves yields compared to traditional episomal methods.
  • The toolkit is effective for both protein overexpression and complex metabolic pathway engineering.