Article Google Scholar 8. The stained cells were observed under a fluorescence microscope. Curr Gene Ther. The vector constructs for homologous recombination were introduced into a transformant in which MTS- Sfo I is expressed under control of the tetracycline Tet -regulated gene expression system to produce heteroplasmic cells with respect to the rps4 gene. Comb for kindly providing p19SfoIRM vector constructs. Conclusion To our knowledge, this is the first report to demonstrate complete and specific disruption of a certain mitochondrial gene by a combination of homologous recombination and introduction of mitochondrially targeted endonuclease. Conclusion The mitochondrial gene-disruption method presented here must be widely useful for precisely determining the functions of individual mitochondrial genes. Please take this quick survey to tell us about what happens after you publish a paper. Figure 3.
Mol Gen Genet.

Apr;(3) The mitochondrial DNA of Dictyostelium discoideum: complete sequence, gene content and genome organization. The Dictyostelium discoideum mitochondrial genome: a primordial system using the universal code and encoding hydrophilic proteins atypical of metazoan.
A third of the total DNA in Dictyostelium is derived from the mitochondrial genome that is present in about copies per cell. Since the 54 kb mitochondrial.
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To reduce this proportion, an effective therapy for these disorders has been tried by use of mitochondrially targeted restriction endonucleases to eliminate specifically mutated mtDNAs [ 671415 ]. This fragment contained the kDa subunit of complex I NADH:ubiquinone oxidoreductaseencoding a predicted amino acid sequence of residues and a molecular mass of 79, daltons which is nuclear encoded in other metazoa.
Figure 1. Additional information Authors' contributions JC performed this research preparing various types of vector constructs.
The Dictyostelium Sequencing Project
The new method presented in this work was devised around the inverse concept. J Cell Sci.
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Thus one can gain rps4 -null cells with only the mutated mtDNA, under the Tet-minus condition.
Strategy for creating rps4 -null cells. We thank P. Video: Dictyostelium mitochondrial genome Mitochondrial DNA inheritance pattern Impairment of mitochondrial function has been associated with a wide range of severe human disorders, which have been regrouped under the name of mitochondrial diseases [ 13 ]. The usefulness of mitochondrially targeted restriction endonucleases in modulating mtDNA is limited by the presence of an appropriate restriction site. Cell Biophys 67—78 Google Scholar. Genomic DNAs were extracted according to the method described previously [ 16 ]. |
This fragment contained the kDa subunit of complex I. Cells that do not contribute genetically to the Dictyostelium zygote nucleus D. discoideum mitochondrial genomes are polymorphic, and our. Abstract. Transcription of the mitochondrial genome in Dictyostelium discoideum gives rise to eight major polycistronic RNA species that can be.
Biol Cell 6: — Google Scholar. Conclusion The mitochondrial gene-disruption method presented here must be widely useful for precisely determining the functions of individual mitochondrial genes.
Additional file 2: Dictyostelium discoideum strains used in this study. Article Google Scholar Bars, 0.

Most of these proteins are encoded in the nucleus, synthesized in the cytosol, and then directed to the mitochondria by specific targeting sequences mitochondria targeting sequence; MTS to be inserted in the mitochondrion by a complex import machinery largely conserved across species.
This seemed to indicate that a majority of Mut-mtDNA was lost during a prolonged time of growth under the Tet-plus condition.
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Genomic Southern hybridization was performed using the Southern method with a slight modification. Staining of cells with a mitochondrion-selective dye, MitoTracker Orange The vegetatively growing cells were incubated in PS medium containing 0.