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RISS 인기검색어
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- 저자 : ( Alicja Ziemienowicz ) (검색결과 2 건)
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( Alicja Ziemienowicz ),( Seyed Mohammad Reza Rahavi ),( Igor Kovalchuk ) 생화학분자생물학회(구 한국생화학분자생물학회) 2011 BMB Reports Vol.44 No.5
The single-stranded DNA binding activity of the Escherichia coli RecA protein is crucial for homologous recombination to occur. This and other biochemical activities of ssDNA binding proteins may be affected by various factors. In this study, we analyzed the effect of CaCl2, NaCl and NH4NO3 salts in combination with the pH and nucleotide cofactor effect on the ssDNA-binding activity of RecA. The studies revealed that, in addition to the inhibitory effect, these salts exert also a stimulatory effect on RecA. These effects occur only under very strict conditions, and the presence or absence and the type of nucleotide cofactor play here a major role. It was observed that in contrast to ATP, ATPγS prevented the inhibitory effect of NaCl and NH4NO3, even at very high salt concentration. These results indicate that ATPγS most likely stabilizes the structure of RecA required for DNA binding, making it resistant to high salt concentrations. [BMB reports 2011; 44(5): 341-346]
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Gene delivery into the plant mitochondria via organelle-specific peptides
Trevor MacMillan,Alicja Ziemienowicz,Fengying Jiang,François Eudes,Igor Kovalchuk 한국식물생명공학회 2019 Plant biotechnology reports Vol.13 No.1
We report a method for genetically engineering the mitochondria of plant cells. Several peptides selected from sorting signal sequences of plant mitochondrial proteins were tested for cell-penetrating, specific organelle-targeting and nucleic acid-binding properties. Selected sequences were named mitochondrial targeting peptides (mTPs). Five of them were used to deliver a linear dsDNA gene construct into AC Ultima spring triticale (X. Triticosecale Wittmack) protoplasts and microspores. The mitochondrial transient expression of the aadA:gfp reporter gene was qualitatively determined with confocal microscopy and quantitatively measured using qRT-PCR. Significant aadA:gfp transcript abundance was reported in protoplasts from deliveries of all five mTPs, while this transcript abundance remained low in microspores. mTP1 transfected microspores were cultured to produce green plantlets and screened for aadA:gfp by endpoint PCR. Targeted gene integration into the mitochondrial genome of eight regenerated haploid triticale lines was confirmed by sequencing. This represents the first report of a successful in vivo transfection of crop plant somatic and germ cells via mitochondrial peptides and a method that can be used to study transient gene expression and to stably deliver genes into the mitochondria of a plant.
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