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Related Concept Videos

Protein Transport to the Stroma01:24

Protein Transport to the Stroma

Chloroplasts are triple membrane structures with an outer membrane, an inner membrane, and a thylakoid membrane, each containing distinct metabolite transporters, membrane translocons, and enzymes. Appropriate sorting and translocating these proteins to their correct membrane systems is essential for chloroplast function.
Protein complexes called the translocon of the outer chloroplast membrane or TOC complex, and the translocon of the inner chloroplast membrane or TIC complex mediate the...
Transgenic Plants02:50

Transgenic Plants

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.
The first-ever transgenic plant was a tobacco plant developed in 1983 that showed resistance against the tobacco mosaic virus. Since then, many transgenic plants have been developed and commercialized for improving the agricultural, ornamental, and horticultural value of a crop plant. Transgenic...
Protein Transport to the Outer Chloroplast Membrane01:11

Protein Transport to the Outer Chloroplast Membrane

Chloroplast outer membrane proteins encoded by the nucleus are synthesized in the cytosol. Soon after synthesis, they bind cytosolic factors such as 14-3-3 protein and the Hsp70 chaperones that keep these precursors in an unfolded state until their translocation.
Two models describe the mechanism of precursor recognition and entry across the outer membrane through the TOC complex. Model 1 suggests the newly synthesized precursor binds to the TOC receptor 159 and forms a complex.
Protein Transport to the Inner Chloroplast Membrane01:18

Protein Transport to the Inner Chloroplast Membrane

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...
Overview of Transposition and Recombination02:13

Overview of Transposition and Recombination

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...
DNA-only Transposons02:57

DNA-only Transposons

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.
The donor site from where the transposon is excised is either degraded or...

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A Simple Method for Isolation of Soybean Protoplasts and Application to Transient Gene Expression Analyses
09:22

A Simple Method for Isolation of Soybean Protoplasts and Application to Transient Gene Expression Analyses

Published on: January 25, 2018

Transient expression in protoplasts.

Fumiaki Katagiri

    CSH Protocols
    |March 2, 2011
    PubMed
    Summary
    This summary is machine-generated.

    Transient gene expression assays offer a quick alternative to studying plant genes. Agrobacterium-mediated leaf assays and root expression are useful for Arabidopsis research.

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    Area of Science:

    • Plant molecular biology
    • Gene expression analysis

    Background:

    • Transient gene expression assays complement studies of stably transformed plants.
    • These assays provide rapid methods for evaluating gene effects using various readouts.
    • Agrobacterium-mediated transformation is a common method for transient assays in plants.

    Purpose of the Study:

    • To evaluate the effectiveness of Agrobacterium-mediated transient gene expression assays in Arabidopsis.
    • To explore alternative transient expression systems in Arabidopsis, such as protoplasts and regenerating roots.
    • To highlight the suitability of root-specific transient expression for investigating root gene function.

    Main Methods:

    • Agrobacterium-mediated transformation of Arabidopsis leaf explants.
    • Transient expression analysis in Arabidopsis protoplasts.
    • Transient expression analysis in regenerating Arabidopsis roots.

    Main Results:

    • Effectiveness of Agrobacterium-mediated leaf assays in Arabidopsis was assessed.
    • Alternative transient expression systems were explored for Arabidopsis.
    • Regenerating roots were identified as suitable for root-specific gene function studies.

    Conclusions:

    • Transient gene expression assays are valuable tools in plant biology research.
    • Agrobacterium-mediated leaf assays and root expression systems are viable options for Arabidopsis.
    • Root-specific transient expression is particularly useful for studying root gene function in Arabidopsis.