Introduction of a leaky stop codon as molecular tool inChlamydomonas reinhardtii
| العنوان: | Introduction of a leaky stop codon as molecular tool inChlamydomonas reinhardtii |
|---|---|
| المؤلفون: | Caspari, Oliver D |
| المصدر: | PLoS ONE PLoS ONE, Vol 15, Iss 8, p e0237405 (2020) |
| بيانات النشر: | Cold Spring Harbor Laboratory, 2020. |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | Chlorophyll, Pigments, 0106 biological sciences, Chloroplasts, Reporter Gene Assay, Chlamydomonas reinhardtii, Plant Science, Biochemistry, 01 natural sciences, Genome, Ribosome, Gene Expression Regulation, Plant, Materials, 0303 health sciences, Multidisciplinary, biology, food and beverages, Genomics, Stop codon, Cell biology, Chloroplast, Physical Sciences, Codon, Terminator, Medicine, Cellular Structures and Organelles, Cellular Types, Research Article, Genetic Markers, Nuclear gene, Imaging Techniques, Science, Plant Cell Biology, Materials Science, Research and Analysis Methods, Genome Complexity, 03 medical and health sciences, Plant Cells, Fluorescence Imaging, Genetics, Molecular Biology Techniques, Molecular Biology, Selectable marker, 030304 developmental biology, Molecular Biology Assays and Analysis Techniques, Reporter gene, Organic Pigments, RuBisCO, Biology and Life Sciences, Computational Biology, Marker Genes, Cell Biology, Ribosomal RNA, biology.organism_classification, Introns, biology.protein, Ribosomes, Selectable Markers, 010606 plant biology & botany |
| الوصف: | Expression of proteins in the chloroplast or mitochondria of the model green algaChlamydomonas reinhardtiican be achieved by directly inserting transgenes into organellar genomes, or through nuclear expression and post-translational import. A number of tools have been developed in the literature for achieving high expression levels from the nuclear genome despite messy genomic integration and widespread silencing of transgenes. Here, recent advances in the field are combined and two systems of bicistronic expression, based on ribosome reinitiation or ribosomal skip induced by a viral 2A sequence, are compared side-by-side. Further, the small subunit of Rubisco (RBCS) was developed as a functional nuclear reporter for successful chloroplast import and restoration of photosynthesis: To be able to combine RBCS with a Venus fluorescent reporter without compromising photosynthetic activity, a leaky stop codon is introduced as a novel molecular tool that allows the simultaneous expression of functional and fluorescently tagged versions of the protein from a single construct. |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::75875af73e4b31c29f3e0e2f2d09e6b1Test https://doi.org/10.1101/2020.04.15.042374Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....75875af73e4b31c29f3e0e2f2d09e6b1 |
| قاعدة البيانات: | OpenAIRE |
| الوصف غير متاح. |