A new method for analyzing gene expression based on frequency analysis of RT-PCR products obtained with degenerate primers

Kiselev, Konstantin V.; Dubrovina, Alexandra S.

doi:10.1007/s11738-009-0426-9

Cited by 26 publications

(8 citation statements)

References 32 publications

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“…The clones were amplified using M13 primers and sequenced, as described previously (Kiselev et al, 2006;Kiselev and Dubrovina, 2010), at the Instrumental Centre of Biotechnology and Gene Engineering of IBSS FEBRAS using an ABI 3130 Genetic Analyzer (Applied Biosystems, Foster City, CA, USA). The BLAST program was used for sequence analysis.…”

Section: Screening Of Actin Pal Dds and Serk Clonesmentioning

confidence: 99%

Mutation of Panax ginseng genes during long-term cultivation of ginseng cell cultures

Kiselev

Shumakova

Tchernoded

2011

Journal of Plant Physiology

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Section: Screening Of Actin Pal Dds and Serk Clonesmentioning

confidence: 99%

Mutation of Panax ginseng genes during long-term cultivation of ginseng cell cultures

Kiselev

Shumakova

Tchernoded

2011

Journal of Plant Physiology

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“…To study gene expression in one non-model plant species like wheat semi-quantitative RT-PCR analysis of total mRNA was chosen because of its exquisite sensibility (Sreenivasulu et al 2007;Hu et al 2009;Kiselev and Dubrovina 2010). Expression of Hsp target genes was assessed with RT-PCR, performed with specific primer pairs.…”

Section: Introductionmentioning

confidence: 99%

Expression of selected heat shock proteins after individually applied and combined drought and heat stress

et al. 2011

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Drought and heat stress are among the abiotic factors causing the most severe damage on plant crops. Their combination is quite common in dry and semi-dry regions worldwide and little is known about its effect on heat shock protein (HSP) profile in wheat plants. The expression of four HSP genes (Hsp 17.8, Hsp 26.3, Hsp 70 and Hsp 101b) in Triticum aestivum L. plants subjected to individually applied water deprivation or high temperature and their combination was monitored via one-step RT-PCR analysis. Changes in the expression levels of small HSPs (smHSPs), HSP70 and HSP100 were established also by SDS-PAGE. The combination of drought and heat induced HSP expression more effectively than the individually applied stresses. The induction of HSPs displayed greater rate in the drought-tolerant wheat variety Katya than in the drought-sensitive cv. Sadovo. The results obtained in wheat plants suggested that the effect of separately applied drought and heat shock cannot be extrapolated to their combination.

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“…PCR products were purified with ethanol and directly sequenced in the Instrumental Centre of Institute of Biology and Soil Sciences (Far Eastern Branch of Russian Academy of Sciences, FEB RAS) using an ABI 3130 Genetic Analyzer (Applied Biosystems) as described previously (Kiselev & Dubrovina 2010, Shumakova et al 2011.…”

Section: Bacterial Strainsmentioning

confidence: 99%

Involvement of the cell-specific pigment genes pks and sult in bacterial defense response of sea urchins Strongylocentrotus intermedius

Kiselev

Ageenko²,

Kurilenko³

2013

Dis. Aquat. Org.

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Bacterial infections are one of the most important problems in mass aquaculture, causing the loss of millions of juvenile organisms. We isolated 22 bacterial strains from the cavity fluid of the sea urchin Strongylocentrotus pallidus and used phylogenetic analysis based on 16S rRNA gene sequences to separate the bacterial strains into 9 genera (Aliivibrio, Bizionia, Colwellia, Olleya, Paenibacillus, Photobacterium, Pseudoalteromonas, Shewanella, and Vibrio). Incubating Strongylocentrotus intermedius larvae with a strain from each of the 9 bacterial genera, we investigated the viability of the larvae, the amount of pigment cells, and the level of polyketide synthase (pks) and sulfotransferase (sult) gene expression. Results of the assay on sea urchin development showed that all bacterial strains, except Pseudoalteromonas and Bizionia, suppressed sea urchin development (resulting in retardation of the embryos' development with cellular disorders) and reduced cell viability. We found that pks expression in the sea urchin larvae after incubation with the bacteria of 9 tested genera was significantly increased, while the sult expression was increased only after the treatment with Pseudoalteromonas and Shewanella. Shikimic acid, which is known to activate the biosynthesis of naphthoquinone pigments, increased the tolerance of the sea urchin embryos to the bacteria. In conclusion, we show that the cell-specific pigment genes pks and sult are involved in the bacterial defense response of sea urchins.

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A new method for analyzing gene expression based on frequency analysis of RT-PCR products obtained with degenerate primers

Cited by 26 publications

References 32 publications

Mutation of Panax ginseng genes during long-term cultivation of ginseng cell cultures

Mutation of Panax ginseng genes during long-term cultivation of ginseng cell cultures

Expression of selected heat shock proteins after individually applied and combined drought and heat stress

Involvement of the cell-specific pigment genes pks and sult in bacterial defense response of sea urchins Strongylocentrotus intermedius

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