• Journal of Plant Biotechnology
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• v.43 no.1
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• pp.37-48
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• 2016

The transcriptional profiles of 'Tamnara' grapevine (Vitis labruscana L.) to Rhizobium vitis were determined using 12,000 gene oligonucleotide microarray chips constructed with 6,776 unigenes based on the EST sequencing. Among them, 95 clones were up-regulated more than three times and 90 were down-regulated more than 5-times in the R. vitis-inoculated grapevines relative to the control vines. Treatment of salicylic acid showed that 337 clones were upregulated and 52 clones were down regulated in grapevines. Microarray analysis, reverse transcription-polymer chain reaction, and slot blot hybridization analysis revealed that 5, 14, and 64 clones were up-regulated and 10, 12, and 61 clones were down-regulated in wounded, salicylic acid-treated, and R. vitis-inoculated 'Tamnara' grapevine leaves, respectively. The expression patterns of ${\beta}$-1,3-glucanase, proline-rich protein, and lipoxygenase genes of 'Tamnara' moderately resistant to R. vitis were similar to those of resistant 'Concord' and 'Delaware' grapevines. However, chalcone synthase genes in 'Tamnara' grapevines showed similar expression patterns to susceptible grapevines 'Neomuscat' and 'Rizamat'. Further expression studies with various clones for each gene should be conducted to elucidate their roles in resistant responses against pathogens or other stimuli in grapevines. These results could provide better resources for understanding the mechanism of defense responses against crown gall disease and clues for identifying new genes that may play a role in defense against R. vitis in grapevines.

• Parasites, Hosts and Diseases
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• v.35 no.3
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• pp.203-210
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• 1997

The difrrrenlial display reverse transcription polymerase chain reaction (DDRT-PCR) aniilysis roils performed to identify the pathogellir strain specific amplicons. mRNAs were purified from the trophozoites of the pathogenif strain YS-27 and the non-pathogenic strain S 16. respectively. Three kinds of rirsl stranded rDNAs were reverse transcribed from the mRNAs by one base anchored oligo-dT 11M (M: A. C, or G) primers. Each cDNA lemplatr was used for DDRT-PCK analysis. A total of 144 pathogenic strain specific amplicons was observed in DDRT-PCR analysis using primer combinations of the 11 arbitrary primers and the 3 one base anchored oli해-dT11M primers. Of these 31 amplit'tons were verified as the amplirons amplified only from the mRNAs of the pathogenic strain by DNA slots biol llybridizatioil. Furthel cklaracleization of the 31 pathogenic strain sprcifil amplicons by DNA slot blot hybridlnation analysis using biotin labeled Probes or the PCR amplified DNA of rysteine proteinase genes revealed that 21 of them were amplliried from the maNAs of the cysteine proteinase genes. Four randomly selected amplirons out of the rest 10 amplirons were used fur screening of cDNA library followed by immunoscreening and all of them were turned outs to be amplified from the mRNA.

• Journal of Microbiology and Biotechnology
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• v.13 no.3
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• pp.385-393
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• 2003

Microbial communities were analyzed in an anaerobic/aerobic sequencing batch reactor (SBR) fed with glucose as a sole carbon source. Scanning electron microscopy (SEM) showed that tetrad or cuboidal packet bacteria dominated the microbial sludge. Quinone, slot hybridization, and 165 rRNA gene sequencing analyses showed that the Proteobacteria beta subclass and the Actinobacteria group were the main microbial species in the SBR sludge. However, according to transmission electron microscopy (TEM), the packet bacteria did not contain polyphosphate granules or glycogen inclusions, but only separate coccus-shaped bacteria contained these, suggesting that coccus-shaped bacteria accumulated polyphosphate directly and the packet bacteria played other role in the enhanced biological phosphorus removal (EBPR). Based on previous reports, the Actinobacteria group and the Proteobacteria beta subclass were very likely responsible for acid formation and polyphosphate accumulation, respectively, and their cooperation achieved the EBPR in the SBR operation which was supplied with glucose.

• The Korean Journal of Zoology
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• v.36 no.4
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• pp.487-495
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• 1993

TRH (Thvrotropin-Releasing Hormone) known to regulate the transcription of the TSH (Thyroid-Stimulating Hormones gene in pituitary cells, but little is understood about the mechanism(sl involved. re present study was attempted to elucidate the role of Ca2+ movement through the voltage-gated channels in the regulation of TSH gene transcription. The c-fos is one of immediate early genes and used as model system for the investigation of signaling pathwavs involved in various stimuli. The changes of c-fos mRNA levels were determined after treatment of various agents using Northern and slot hybridization analysis. The c-fos mRNA was rapidly and transiently induced by TRH (about 3-fold) in GH3 cells and this induction was repressed by calcium chelating agent (EGTA), calcium channel blocker (verapamil) anti protein kinase C inhibitor (aminoacridine). The abilities of forskolin (adenvlate cvclase activators, PMA (protein kinase C activator), and A23187 (calcium ionophore) to affect c-ios gene transcription, either alone or in combination with TRH were tested in the same cells. All of them significantly increased the level of c-fos mRUA. However, no additive relationship was observed in all combined treatments except forskolin. These results suggest that TRH action on the c-fos gene activation is mediated by calcium influx as well as through protein kinase C.

• Journal of Microbiology and Biotechnology
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• v.18 no.7
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• pp.1290-1297
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• 2008

To investigate the diversities of Accumulibacter phosphatis and its polyhydroxyalkanoate (PHA) synthase gene (phaC) in enhanced biological phosphorus removal (EBPR) sludge, an acetate-fed sequencing batch reactor was operated. Analysis of microbial communities using fluorescence in situ hybridization and 16S rRNA gene clone libraries showed that the population of Accumulibacter phosphatis in the EBPR sludge comprised more than 50% of total bacteria, and was clearly divided into two subgroups with about 97.5% sequence identity of the 16S rRNA genes. PAO phaC primers targeting the phaC genes of Accumulibacter phosphatis were designed and applied to retrieve fragments of putative phaC homologs of Accumulibacter phosphatis from EBPR sludge. PAO phaC primers targeting $G_{1PAO},\;G_{2PAO},\;and\;G_{3PAO}$ groups produced PCR amplicons successfully; the resulting sequences of the phaC gene homologs were diverse, and were distantly related to metagenomic phaC sequences of Accumulibacter phosphatis with 75-98% DNA sequence identities. Degenerate NPAO (non-PAO) phaC primers targeting phaC genes of non-Accumulibacter phosphatis bacteria were also designed and applied to the EBPR sludge. Twenty-four phaC homologs retrieved from NPAO phaC primers were different from the phaC gene homologs derived from Accumulibacter phosphatis, which suggests that the PAO phaC primers were specific for the amplification of phaC gene homologs of Accumulibacter phosphatis, and the putative phaC gene homologs by PAO phaC primers were derived from Accumulibacter phosphatis in the EBPR sludge. Among 24 phaC homologs, a phaC homolog (GINPAO-2), which was dominant in the NPAO phaC clone library, showed the strongest signal in slot hybridization and shared approximately 60% nucleotide identity with the $G_{4PAO}$ group of Accumulibacter phosphatis, which suggests that GINPAO-2 might be derived from Accumulibacter phosphatis. In conclusion, analyses of the 16S rRNA and phaC genes showed that Accumulibacter phosphatis might be phylogenetically and metabolically diverse.

• Preventive Nutrition and Food Science
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• v.5 no.3
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• pp.153-159
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• 2000

A 4.4 kb DNA fragment encompassing lacA (galactoside acetyltransferase) and lacZ($\beta$-galactosidase) genes from Lactococus lactis ssp. lactis ATCC 7962 (L. lactis 7962) was introduced ito a Lac strain, Lactococcus lactis ssp. lactis MG1363 (L. lactis MG1363) by using a lactococcal expression vector, pMG36e and expression level of lacZ was examined. Growth rates and $\beta$-galactosidase ($\beta$-gal) activities of MG1363 cells carrying recombinant plasmid, pMLZ3, on M17 broth containing different carbon sources (1%, w/v) were examined. Contrary to the expectations, MG1363 [pMLZ3] grown on lactose showed the lowest enzyme activity (17 units) and cells grown on galactose had the highest $\beta$-gal activity (41 units). Cells grown on glucose had intermediate activity (33 units). These activities are about one tenth of the values observed in L. lactis 7962 where lacZ is present as a single-copy gene in the chromosome. When the cellular concentrations of lacZ transcript were examined using slot blot hybridization, it was found that MG1363[pMLZ3] produced sufficient amounts of transcript. These results indicate that either proteolytic degradation of $\beta$-gal or other regulatory mechanism prevent the translation or accumulation of $\beta$-gal in L. lactis MG1363 cells. In regard to regulation, the presence of the ccpA gene in L. lactis MG1363 was confirmed by Southern blot.

• Journal of Microbiology and Biotechnology
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• v.9 no.2
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• pp.201-205
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• 1999

The effects of growth temperature and a carbon source on the expression of $\beta$-galactosidase gene of Lactococcus lactis ssp. lactis ATCC 7962 (L. lactis 7962) were investigated. At $25^{\circ}C$, L. lactis 7962 had a higher $\beta$-galactosidase activity than cells grown at $30^{\circ}C$ or $37^{\circ}C$, although cells grew most quickly at $37^{\circ}C$ The highest $\beta$-galactosidase activity was observed in cells grown in M17 with lactose (l %) followed by cells grown in a galactose (1 %) medium. L. lactis 7962 exhibited the minimum $\beta$-galactosidase activity in glucose media, indicating catabolite repression. When the cellular levels of $\beta$-galactosidase mRNA were examined using slot blot hybridization, no significant differences were observed between cells grown at $25^{\circ}C$ and cells at $30^{\circ}C$ or $37^{\circ}C$ in the same media. This suggests that the quantity of $\beta$-galactosidase mRNA may not be the reason for the higher $\beta$-galactosidase activities of L. lactis 7962 at $25^{\circ}C$ The level of ccpA (Catabolite Control Protein) transcript remained almost constant during the exponential growth phase irrespective of a carbon sourse.

• The Plant Pathology Journal
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• v.15 no.5
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• pp.295-301
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• 1999

Many of plant defense responses are consequence of transcriptional activation of related genes. We have developed a modified differential screening procedure to isolate tobacco genes that are involved in the defense responses against TMV infection. A cDNA library was constructed from Nicotiana glutinosa leaves infected by TMV under temperature shift conditions. Each of plasmid DNA in the library was hybridized on a set of slot blots to a pool of cDNA probes prepared from either TMV-infected or mock-treated tobacco leaves. Among 900 plasmid DNAs, 81 clones exhibiting significantly enhanced or reduced level of hybridization to either probe were selected for nucleotide sequencing. The clones were listed into 61 genes considering redundancy between the sequences. The genes were identified to be defense-related genes including PR-genes and genes involved in primary or secondary metabolisms. This results supports the implication that plant defense process entails a major shift in total cellular metabolisms rather than activation of a limited number of defense-related genes. Expression patterns of a number of defense-related genes. Expression patterns of a number of selected genes were examined in northern blot analyses. It is notable that the clone 630 of unknown function exhibits expression pattern similar to those of previously known PR-genes. Experiments to elucidate the roles in defense mechanism of a couple of genes newly identified in this study are in progress.

• Proceedings of the Ginseng society Conference
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• 1990.06a
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• pp.58-64
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• 1990

The effects of ginseng saponins, G-Rbl and G-Rc on the rat liver LDH A-gene transcnptional activity was investigated during pro-replicative phase of rat liver after partial hepatectomy. Changes in LDH A-mRNA levels in regenerating rat liver after intraperitoneal administrations of G-Rbl of G-Rc were tested by slot blot hybridization methods. The results showed that G-Rbl (1 mg/100g B.W) and G-Rc (1 ma/100g B.W) caused marked increases of LDH A-mRNA contents by respectively 1.9- and 1.5-fold in rat liver at 5·hours after partial hepatectomy. Dose dependent effect of G-Rbl and G-Rc (1-25 mg/100g B.W) on the LDH A-mRNA levels on regenerating rat liver were also analyzed. The maximal in- creases of liver LDH A-mRNA levels were observed with the doses of 1 mg for G-Rbl and 5 mg for G-Rc However, when the administration doses of G-Kbl and G-Rc were increased to 20 mg, G-Rbl caused a marked decrease of LDH A-mRNA level to 61% of those in sham-operated rat liver In contrast, G-Rc slightly decreased the liver LDH A-mRNA contents by 30% as compared to those of the maximum value but still maintained 22% higher LDH A-mRNA levels then those of sham-operated rate liver. On the basis of these experimental results, we conclude that ginseng saponin, G-Rb 1 and G·Rc have stimulatory effect at the lower concentration (1 mg/100g B.W) and inhibitory effect at the higher concentration (20 moi loos 5.W) on the LDH A-gene transcription during regeneration of rat liver, Additionally we also investigated the stimulatory effects of ginsenosides on the protein and DNA synthetic activities in hepatocyte primary cell cultures isolated from regenerating rat liver. Both of G·Rc and -Re increased the synthetic rates of hepatocytes proteins and DNA at the administration doses of 50 ug and 100 ug/3 ml/dish respectively representing 1.3-1.6 fold increases. From these results we postulate that G-Rc and -Re may have a mitogen enhancer activity for the hepatocyte proliferation during rat liver regeneration period. Keywords Inductive effects of ginsenosides, G-Rb, -Rc, and -Re, rat LDH A-gene transcription, the sin thetic rate of proteins and DNA in regeneration rat liver.

• Journal of Ginseng Research
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• v.14 no.2
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• pp.200-206
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• 1990

The effects of ginseng saponins, SRbl and G-Rc on the rat liver LDH A-gene transcriptional activity was investigated during prereplicative phase of rat liver after partial hepatectomy. Changes in LDH A-mRNA levels in regenerating rat liver after intraperitoneal administrations of G-Rbl or 'G-Rc were tested by slot blot hybridization methods. The results showed that G-Rbl (1 mg/100g B.W) and G-Rc (1 mg/100g B.W) caused marked increases of LDH A-mRNA contents by respectively 1.9- and 1.5-fold in rat liver at 5-hours after partial hepatectomy Dose dependent elect of G-Rbl and G-Rc (1-25 mg/ 100g B.W) on the LDH A-mRNA levels on regenerating rat liver were also analyzed. The maximal increases of liver LDH A-mRNA levels were observed with the doses of 1 mg for G-Rbl and 5 mg for G-Rc. However, when the administration doses of G-Rbl and G-Rc were increased to 20 mg, G-Rbl caused a marked decrease of LDH A-mRNA level to 61% of those in sham-operated rat liver. In contrast, G-Rc slightly decreased the liver LDH A-mRNA contents by 30% as compared to those of the maximum value but still maintained 22% higher LDH A-mRNA levels then those of sham-operated rate liver. On the basis of these experimental results, we conclude that ginseng saponin, G-Rbl and G-Rc have stimulatory effect at the lower concentration (1 mg/ 100g B.W) and inhibitory effect at the higher concentration (20 mg/ 100g B.W) on the LDH A-gene transcription during regeneration of rat liver. Additionally we also investigated the stimulatory effects of ginsenosides on the protein and DNA sinthetic activities in hepatocyte primary cell cultures isolated from regenerating rat liver. Both of G-Rc and -Re increased the synthetic rates of hepatocytes proteins and DNA at the administration doses of 50 us and 100 $\mu\textrm{g}$/3 ml/dish respectively representing 1.3-1.6 fold increases. From these results we postulate that G-Rc and -Re may have a mitogen ehincer activity for the hepatocyte proliferation during rat liver regeneration period.