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  • Chlamydomonas  (1)
  • RNA processing  (1)
  • Springer  (2)
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  • Springer  (2)
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    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 256 (1997), S. 88-91 
    ISSN: 1617-4623
    Keywords: Key wordsSaccharomycescerevisiae ; Mitochondria ; COX1 ; RNA processing ; RT-PCR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Splicing of the group I intron aI5β from the yeast mitochondrial COX1 transcript requires at least four proteins, encoded by the nuclear genes PET54, MRS1/PET157, SUV3 and MSS18. These proteins either act directly to facilitate intron aI5β excision, or indirectly in some manner. One possible indirect mode of action of these nuclear gene products is in stimulation of expression of a mitochondrial protein, such as a maturase, that is necessary for intron aI5β excision. To test this possibility, splicing of intron aI5β was examined in a ρ− strain, which is incapable of mitochondrial protein synthesis. A quantitative RT-PCR assay was set up to compare levels of spliced COX1 mRNA present in three strains: a wild–type ρ+ strain; the ρ−strain 7–49b-11, which retains the entire COX1 transcription unit; and a strain bearing a null mutation in the nuclear PET54 gene. The results showed that excision of aI5β occurs relatively efficiently in the ρ− strain, and therefore does not require any mitochondrial-encoded proteins.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1617-4623
    Keywords: Key words Bioluminescence ; Chlamydomonas ; Chloroplast transformation ; Chloroplast translation ; Vital reporter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The use of luciferases as reporters of gene expression in living cells has been extended to the chloroplast genome. We show that the luciferase from the soft coral Renilla reniformis (Rluc) can be successfully expressed in the chloroplast of Chlamydomonas reinhardtii. Expression of the rluc cDNA was driven by the promoter and 5′ untranslated regions of the atpA gene. Western analysis with an anti-Rluc antibody detected a single polypeptide of 38 kDa in the luminescent cells. This is 3 kDa larger than native Rluc, and suggests that translation of the chimeric mRNA begins at the atpA start codon, 29 codons upstream from the rluc start site. We also show that the luminescence of the transformants was sufficient to enable imaging of colonies using a cooled CCD camera.
    Type of Medium: Electronic Resource
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