• Applied Biological Chemistry
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• v.42 no.2
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• pp.123-127
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• 1999

Iron is an essential nutrient but potentially toxic element in human. To identify the effects of iron on the gene expression of mammalian cell, we have isolated several genes that are regulated by iron using the RNA differential display method. RNAs were isolated from HeLa cells treated with iron supplement or iron chelator. A total of 24 genes were isolated and of these, four genes were identified by DNA sequencing and northern blot.

• Journal of Plant Biology
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• v.35 no.3
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• pp.253-257
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• 1992

Potyvirus group is the largest group among plant virus groups and damages severely plant hosts upon infectiQn. In order to investigate the mechanism by which potyviruses induce disease in plants, a cDNA clone 29-6 which is cOIlsidered to be a cDNA clone for garlic mosaic virus (GMV) was isolated. It did not hybridize to garlic latent virus genome, which is one of two major garlic viruses. Northern blot analysis shows that the genome size of garlic mosaic virus was about 9 kb. Clone 29-6 strongly hybridizes to poly(A) RNA isolated from garlic leaves, suggesting that GMV RNA is polyadenylated as other potyviruses. Nucleotide sequence analysis of cDNA clones overlapping with clone 29-6 showed that garlic plants are infected with various strains of garlic mosaic virus which are closely related to each other. other.

• Korean Journal of Plant Resources
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• v.10 no.4
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• pp.300-304
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• 1997

Leaf dise explants of Solanum tuberosum cultivar. Desiree and Atlantic, were infected with a Agrobacterium MP90 strain containing chimeric gene construct, consisting of antibiotic and low temperature regulated gene (H28) for transformation. regenerated multiple shoots were selected on a medium containing kanamycin and carbenieillin after exposure to Agrobacterium. Both PCR analysis of NPT Ⅱ, H28 genes and northern blot analysis indicated that the genes coding for the enzyme were successfully integrated into the potato genome and could be expressed in potato plants.

• Animal cells and systems
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• v.9 no.4
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• pp.205-209
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• 2005

A rapid, simple and efficient method to extract RNA from the adult polyps of a soft coral, marine cnidarian, Scleronephthya gracillimum $(K\ddot{u}kenthal)$; was developed in this study. The highest yield and purity of RNA was obtained with the lysis solution containing 35 mM EDTA, 0.7 M LiCl, 7.0% SDS, and 200 mM Tris-Cl (pH 9.0). Approximately $40{\mu}g$ of total RNA was extracted from 200 mg of liquid nitrogen-pulverized polyp tissue. The ratio of absorbance at 260 nm and 280 nm ranged from 1.8 to 2.0. The results of the reverse transcription polymerase chain reaction (RTPCR) with ${\beta}-actin$ gene specific primers and Northern blot analysis using the same gene probe revealed that the RNA extracted by our method had high quality, and was sufficient for subsequent molecular biological analyses. This method was effective for RNA extraction from other soft coral species which belong to the genus Dendronephthya.

• Archives of Pharmacal Research
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• v.22 no.6
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• pp.542-548
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• 1999

The UL47 gene (b 60390-b 60388) located in the unique long region of the human cytomegalovirus (HCMV) AD169 strain genome was analyzed RNA mapping. Northern blot analysis showed that the UL47 gene was expressed at late times after infection (72 h postinfection). The 9.7-kb transcript was expressed in the infected cells but not in phosphonoformate-treated cells at 72 hpi, indicating that the UL47 gene was only expressed at late times after infection. To map the 5'-end and 3'-end of UL47 transcripts, primer at late times after infection. To map the 5'-end and 3'-end of UL47 transcripts, primer extension and RNase protection analysis were performed. Primer extension analysis revealed that the transcription initiation site of UL47 was located in 27 bp downstream (b 60323) of the TATA box motif. The sizes of UL47 ORF (approximately 2.9-kb) and UL48 ORF (approximately 6.7-kb) deduced from computer sequence analysis suggest that the expressed 9.7-kb transcript of UL47 uses the 3'-end polyadenylation signal of Ul48. The result of RNase protection determined that the 3'-end of UL47 RNA utilized the 3'-end polyadenylation signal of UL48, which is located in HCMV genome b 70082.

• Korean Journal of Microbiology
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• v.34 no.4
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• pp.243-247
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• 1998

Comparative analyses of intracellular biopterin contents and its biosynthetic enzymes were performed in Allomyces macrogynus. Biopterin content in fresh weight was 14-fold higher in mycelium than in zoospore. Enzyme activities of GTP cyclohydrolase I and 6-pyruvoyltetrahydropterin synthase in ammonium sulfate fractions were approximately 2-fold higher in mycelium. On the other hand, sepiapterin reductase (SR) activity was 10 fold higher in zoospore. Northern blot assay also demonstrated that SR transcript was abundant in zoospore. These results suggest a possible involvement of tetrahydrobiopterin in cellular differentiation of Allomyces macrogyllus as well as provide an experimental basis to elucidate the physiological function of SR in this organism.

• Korean Journal of Plant Tissue Culture
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• v.22 no.4
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• pp.195-200
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• 1995

The mammalian adenosine deaminase(ADA) gene was stably expressed in transgenic tobacco plane. The chimeric ADA gene 35S/35S/AMV/ADA/Tnos, has been constructed. This chimeric gene was introduced into the binary vector pRD400, which was thereafter mobilized into Agrobacterium tumefaiens strain MP90 harboring disarmed Ti-plasmid. The resulting strains were used to transform Nicofiana tabacum L. using the leaf disc. Incorporation of the chimeric gene into plant were confirmed by PCR and Northern blot analyses. Immunoblot analysis showed that ADA protein was successfully synthesized in the transgenic tobacco plants.

• Molecules and Cells
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• v.42 no.4
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• pp.356-362
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• 2019

The binding of MS2 bacteriophage coat protein (MCP) to MS2 binding site (MBS) RNA stem-loop sequences has been widely used to label mRNA for live-cell imaging at single-molecule resolution. However, concerns have been raised recently from studies with budding yeast showing aberrant mRNA metabolism following the MS2-GFP labeling. To investigate the degradation pattern of MS2-GFP-labeled mRNA in mammalian cells and tissues, we used Northern blot analysis of ${\beta}$-actin mRNA extracted from the Actb-MBS knock-in and $MBS{\times}MCP$ hybrid mouse models. In the immortalized mouse embryonic cell lines and various organ tissues derived from the mouse models, we found no noticeable accumulation of decay products of ${\beta}$-actin mRNA compared with the wild-type mice. Our results suggest that accumulation of MBS RNA decay fragments does not always happen depending on the mRNA species and the model organisms used.

• Korean Journal of Plant Tissue Culture
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• v.25 no.5
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• pp.341-346
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• 1998

In this paper we describe the cloning and expression of the genes encoding the flavonoid-biosynthetic enzyme dihydroflavonol 4-reductase (DFR) in Matthiola incana R. Br. A heterologous cDNA probe from Zea mays was used to isolate full-size DFR cDNA clone from a corolla-specific cDNA library. Comparison of the coding region of this DFR cDNA sequence including the sequences of Zea mays, Anthirrinum majus, Petunia hybrida, Callistephus chinensis, Dianthus caryophyllus and Rosa hybrida reveals a identity higher than 61% at the nucleotide level. The DFR transcript is G/C rich in monocotyledonous plants show a strong codon bias preferring codons with a G or C in the third position. The function of this nucleotide sequences were verified by comparison with amino acid sequences of the amino-terminus and tryptic peptides from purified plant enzyme, by northern blotting with mRNA from wild type and mutant plants and by in vitro expression yielding an enzymatically active reductase. Genomic southern blot analysis showed the presence of one gene for DFR in Matthiola incana. Northern blot analysis of the DFR wild type and mutant lines showed that the lack of DFR activity in the stable acyanic mutant k17b is clearly by a transcriptional block of the DFR gene.

• Microbiology and Biotechnology Letters
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• v.30 no.1
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• pp.15-20
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• 2002

Diacylglycerol kinase (DGK) phosphorylates the second messenger diacylglycerol (DAG) to phosphatidic acid and it may play a role in signal transduction in Escherichia coli as well as in eukaryotic cells. In addition, DGK is important for microorganisms to adapt to several physiological stimuli. In Bacillus subtilis, the effect of stress on dgk transcription was examined by northern hybridization. The high level of dgk transcription was induced against high osmolarity, low pH value and low temperature. Transcriptional analysis revealed that the dgk gene and dgk upstream locus (ORF2, ORF3 and ORF4) were transcribed as a polycistronic mRNA to form an approximately 2.5 kb transcript.