Fluorescent in situ hybridization of mitochondrial DNA and RNA. | Zendy

Lukáš Alán | Zendy; Jaroslav Zelenka | Zendy; Jan Ježek | Zendy; Andrea Dlasková | Zendy; Petr Ježek | Zendy

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Fluorescent in situ hybridization of mitochondrial DNA and RNA.

Author(s) -

Lukáš Alán,

Jaroslav Zelenka,

Jan Ježek,

Andrea Dlasková,

Petr Ježek

Publication year - 2010

Publication title -

acta biochimica polonica

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.452

H-Index - 78

eISSN - 1734-154X

pISSN - 0001-527X

DOI - 10.18388/abp.2010_2425

Subject(s) - mitochondrial dna , nucleoid , molecular beacon , heavy strand , microbiology and biotechnology , dna , transcription bubble , biology , nucleic acid , rna , mt rnr1 , fluorescence , rnase p , gene , biophysics , genetics , rna polymerase , oligonucleotide , physics , optics , transfer rna , escherichia coli

To reveal nucleic acid localization in mitochondria, we designed molecular beacon fluorescent probes against: i) the light strand complementary to ND5 mitochondrial DNA (mtDNA) gene (annealing also to corresponding mRNA); ii) displacement (D) loop 7S DNA (annealing also to parallel heavy strand mtDNA and corresponding light strand transcript); iii) the proximal D-loop heavy strand displaced by the light strand promoter minor RNA. Confocal microscopy demonstrated ND5 probe spreading (less for other probes) in mitochondrial reticulum tubules but upon RNase A treatment all probes contoured mtDNA nucleoid localization. DNase I spread the signal over mitochondrial tubules. Future applications are discussed.

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