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A eukaryotic cell can have up to three different types of genetic systems: nuclear, mitochondrial, and chloroplast. During evolution, organelles have exported many genes to the nucleus; this transfer is still ongoing in some plant species. Approximately 18% of the Arabidopsis thaliana nuclear genome is thought to be derived from the chloroplast’s cyanobacterial ancestor, and around 75% of the yeast genome derived from the mitochondria’s bacterial ancestor. This export has occurred...
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The present-day mitochondrial and chloroplast genomes have retained some of the characteristics of their ancestral prokaryotes and also have acquired new attributes during their evolution within eukaryotic cells. Like prokaryotic genomes, mitochondrial and chloroplast genomes neither bind with histone-like proteins nor show complex packaging into chromosome-like structures, as observed in eukaryotes. Unlike mitotic cell divisions observed in eukaryotic cells, mitochondria and chloroplasts...
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Recombinant DNA technology called transgenesis is often used to add a foreign gene or remove a detrimental gene from an organism. Such genetically modified organisms are called transgenic organisms.
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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.
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Plasmids

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Plasmids are extrachromosomal DNA molecules found in bacteria, archaea, and some eukaryotic microbes like yeast. These small, circular DNA structures typically contain fewer than 30 genes, although some may exist linearly. Plasmids vary in their number within a cell, known as copy number. Single-copy plasmids are present in one copy per cell and multi-copy plasmids are present in multiple copies, reaching over 100 copies per cell.Plasmids usually replicate independently of the chromosomal DNA...
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Proteins targeted to the inner chloroplast membrane, or plastid proteins, are transported by two general pathways: the stop-transfer and the re-insertion or post-import pathways. Most plastid proteins carry N-terminal transit sequences and internal import sequences targeting it to the specific chloroplast subcompartment. Proteins targeted by the stop-transfer pathway have internal hydrophobic sequences that inhibit their translocation into the stroma. As a result, these precursors are arrested...
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Co-expression of Multiple Chimeric Fluorescent Fusion Proteins in an Efficient Way in Plants
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Mini-synplastomes for plastid genetic engineering.

Alessandro Occhialini1,2, Alexander C Pfotenhauer1,2, Li Li1,2

  • 1Department of Food Science, University of Tennessee, Knoxville, TN, USA.

Plant Biotechnology Journal
|September 29, 2021
PubMed
Summary
This summary is machine-generated.

Scientists developed a novel

Keywords:
Solanum tuberosumepisomal replicationhomologous recombinationplastid engineeringplastomesmall synthetic plastome ‘mini-synplastome’

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Area of Science:

  • Plant Biotechnology
  • Synthetic Biology
  • Molecular Biology

Background:

  • Plastid engineering is crucial for synthetic biology applications in plants.
  • Current methods like homologous recombination are limited to small constructs and are over 30 years old.
  • A nimble platform is needed for introducing novel synthetic circuits into plant plastids.

Purpose of the Study:

  • To design, build, and test a novel synthetic genome structure for plant plastids.
  • To create a system that allows for facile, marker-free plastid engineering.
  • To establish a platform for one-step metabolic engineering in plants.

Main Methods:

  • Inspired by dinoflagellate plastome organization, a 'mini-synplastome' was designed.
  • The mini-synplastome was developed in vitro to meet specific criteria.
  • Key criteria included episomal replication, facile cloning, predictable expression, non-integration, and autonomous persistence.

Main Results:

  • The first plant mini-synplastome was successfully developed in vitro.
  • The developed mini-synplastome meets all specified design criteria.
  • This novel structure allows for episomal replication and autonomous persistence without integrating into the native plastome.

Conclusions:

  • The mini-synplastome represents a revolutionary advancement in chloroplast biotechnology.
  • This platform enables facile, marker-free plastid engineering.
  • It offers an unparalleled system for one-step metabolic engineering in plants.