• Microbiology and Biotechnology Letters
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• v.35 no.3
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• pp.177-183
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• 2007

Using complementation of RpoH deficient E. coli strain A7448, the rpoH gene encoding heat shock sigma factor 32 (${\sigma}^{32}$) from Methylovorus sp. strain SS1 DSM11726 was cloned and sequenced. Sequence analysis of a stretch of 1,796-bp revealed existence of an open reading frame encoding a polypeptide of 284 amino acid (32,006 dalton). Deduced amino acid sequence of the Methylovorus sp. strain SS1 RpoH showed that 59.6%, 39.1% and 51.4% identities with those of Nitrosomonas europaea (${\beta}$-proteobacteria), Agrobacterium tumefaciens ($\alpha$-proteobacteria) and E. coli (${\gamma}$-proteobacteria). The expression level of the functional ortholog of RpoH of Methylovorus sp. strain SS1 was increased transiently after heat induction, further indicating that it functions as a heat shock sigma factor.

• Korean Journal of Medicinal Crop Science
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• v.22 no.6
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• pp.489-496
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• 2014

Polygonatum is a genus placed in the family Liliaceae, distributed throughout the Northern Hemisphere and 16 of the species are grown naturally in Korea. In oriental medicine, the rhizomes of Polygonatum have been used as two different medicines, Okjuk (Polygonati odorati Rhizoma) and Hwangjeong (Polygonati Rhizoma). However, it is difficult to identify the morphological and chemical differences between the medicinal groups and thus easy to confuse the one with the other. Therefore, a clear classification standard needs to be established so as to be able to discriminate between them. In the study, the morphological characteristics of the plants, Polygonatum spp., were examined. Then, the differences in SNPs among the DNA sequences of 7 of the Polygonatum spp. and 1 of the Disporum spp. were analyzed by DNA barcoding with rpoC1, rpoB2, matK, and psbA-trnH of the cpDNA region. In the results, three regions, rpoC1, rpoB2, and matK were useful for discriminating the species, P. stenophyllum and P. sibiricum. Furthermore, it was possible to discriminate the individual germplasm within the species by using the combination of the results obtained from rpoB2, rpoC1, and matK.

• Journal of Pharmacopuncture
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• v.15 no.2
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• pp.20-23
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• 2012

Panax ginseng is a well-known herbal medicine in traditional Asian medicine, and wild ginseng is widely accepted to be more active than cultivated ginseng in chemoprevention. However, little has actually been reported on the difference between wild ginseng and cultivated ginseng. Thus, to identify and analyze those differences, we used suppressive subtraction hybridization (SSH) sequences with microarrays, realtime polymerase chain reaction (PCR), and reverse transcription PCRs (RT-PCRs). One of the clones isolated in this research was the chloroplast rpoC1 gene, a ${\beta}$subunit of RNA polymerase. Real-time RT-PCR results showed that the expression of the rpoC1 gene was significantly upregulated in wild ginseng as compared to cultivated ginseng, so, we conclude that the rpoC1 gene may be one of the important markers of wild ginseng.

• Journal of Microbiology and Biotechnology
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• v.29 no.8
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• pp.1299-1309
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• 2019

As an opportunistic bacterial pathogen, Pseudomonas aeruginosa PAO1 contains two phenazine-producing gene operons, phzA1B1C1D1E1F1G1 (phz1) and phzA2B2C2D2E2F2G2 (phz2), each of which is independently capable of encoding all enzymes for biosynthesizing phenazines, including phenazine-1-carboxylic acid and its derivatives. Other previous study reported that the RpoS-deficient mutant SS24 overproduced pyocyanin, a derivative of phenazine-1-carboxylic acid. However, it is not known how RpoS mediates the expression of two phz operons and regulates pyocyanin biosynthesis in detail. In this study, with deletion of the rpoS gene in the $PA{\Delta}phz1$ mutant and the $PA{\Delta}phz2$ mutant respectively, we demonstrated that RpoS exerted opposite regulatory roles on the expression of the phz1and phz2 operons. We also confirmed that the phz1 operon played a critical role and especially biosynthesized much more phenazines than the phz2 operon when the rpoS gene was knocked out in P. aeruginosa. By constructing the translational reporter fusion vector lasR'-'lacZ and the chromosomal fusion mutant $PA{\Delta}lasR::lacZ$, we verified that RpoS deficiency caused increased expression of lasR, a transcription regulator gene in a first quorum sensing system (las) that activates overexpression of the phz1 operon, suggesting that in the absence of RpoS, LasR might act as an intermediate in overproduction of phenazine biosynthesis mediated by the phz1 operon in P. aeruginosa.

• Korean Journal of Microbiology
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• v.37 no.3
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• pp.228-233
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• 2001

The acid tolerance response (ATR) of log-phase Salmouella enterica seroyar Typhimurium is induced by acid adaptation below pH4.5 and will protect cells against more severe acid. Two distinctive ATR systems in thisorganism are a log-phase and stationary-phase ATR in which acid adaptations trigger the synthesis of acid shockproteins (ASPs). We found that log-phase ATR system was strongly affected by environmental factor, low tem-perature, $25^{\circ}C$. Exposure to low temperature and mild acid has been shown to increase acid survival dra-matically, and this survival rate was showed higher than $37^{\circ}C$. Especially unadapted cells at $25^{\circ}C$ presented tenthousand folds survival increasing when compared with cells at $37^{\circ}C$. The degree of acid tolerance of rpoSwhich is blown to be required for acid tolerance more increase than $37^{\circ}C$. Even though AIR pattern of rpoSbetween unadapted and adapted was showed similar at pH 3.1, rpoS-dependent ATR system also has beendetected in low temperature because rpoSAp prevents sustained acid survival at $25^{\circ}C$. Therefore the resultssuggest low temperature ATR system requires rpoS-dependent and -independent both. To investigate the basisfor low temperature related ATR system, gene that was participated for low temperature acid tolerance (lat) wasscreened in virulent S. enterica serovar Typhimurium UKl Using the technique of P22- MudJ (Km, lacZ)-directed lacZ operon fusion, LF452 latA‥‥MudJ was isolated. The latA‥‥MudJ of S. enterica Typhimurium pre-vented low temperature acid tolerance response. Therefore latA is considered one of the important genes for acidadaptation.

• Journal of Marine Bioscience and Biotechnology
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• v.2 no.4
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• pp.263-266
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• 2007

Vibrio vulnificusis a causative agent of serious diseases in humans resulting from the contact of wound with seawater or consumption of raw seafood. Several studies aimed at detecting V. vulnificus have targeted vvh as a representative virulence toxin gene belonging to the bacterium. In this study, we targeted the rpoS gene, a general stress regulator, to detect V. vulnificus. PCR specificity was identified by amplification of 8 V. vulnificus templates and by the loss of a PCR product with 36 non-V. vulnificus strains. The PCR assay had the 273-bp fragment and the sensitivity of 10 pg DNA from V. vulnificus. SYBR Green I-based real-time PCR assay targeting the rpoS gene showed a melting temperature of approximately $84^{\circ}C$ for V. vulnificus strains. The minimum level of detection by real-time PCR was 2 pg of purified genomic DNA, or $10^3$ V. vulnificus cells from pure cultured broth and $10^3$ cells in 1g of oyster tissue homogenates. These data indicate that real-time PCR is a sensitive, species-specific, and rapid method for detecting this bacterium using the rpoS gene in pure cultures and in infected oyster tissues.

• Korean Journal of Microbiology
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• v.32 no.4
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• pp.264-270
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• 1994

A gene coding for a novel putative $\sigma$ factor of RNA polymerase has been identified from Streptomyces coelicolor A3(2) using Escherichia coli rpoS gene fragment as a probe. The 486 bp rpoS gene fragment was amplified from E. coli genomic DNA by PCR with two synthetic oligonucleotides, the sequences of which were deduced from the amino acid sequences in the regions 2.3 and 4.2 conserved among various bacterial factors. When E. coli genomic DNA fragments were hybridized with cloned rpoS probe, only one band corresponding to rpoS gene (3.2 kb PvuII fragment or 2.3 kb KpnI fragment) was detected. In S. coelicolor, however, two bands were detected both in PvuII digested DNA and SalI digested DNA. 3.5 kb PvuII fragment which binds the rpoS gene probe was cloned (pMS1) from the sublibrary, and the nucleotide sequences of 1.0 kb BamH'/HincII subclone (pBH2) was partially determined. The nucleotide sequences revealed extensive similarity to other $\sigma$ factor genes of S. coelicolor (hrdA, hrdB, hrdC, hrdD), S. aureofaciens (hrdA, hrdB, hrdC, hrdD), Synechococcus species, Pseudomonas aeruginosa, Stigmatella aurantiaca, and Anabaena species. The nucleotide sequences in regions 1.2 and 4 were compared with the corresponding regions of 5 known ${\sigma}$ factor genes of S. coelicolor by multiple alignment. It turned out that the cloned gene is most closely related to hrdA showing 88% amino acid similarity in region 1.2 and 75% in region 4.

• Archives of Pharmacal Research
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• v.18 no.5
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• pp.336-339
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• 1995

Ephedra distachya cultures have been known to accumulate two major $\rho-coumaroylamino$ acids (p-coumaroylgicine and $\rho-coumaroylamino$ -D-alanine) by D-Ala treament. When D-Ala was added together with serial concentrations of yeast-derived elicitor, the accumulation of $\rho-coumaroylamino$ -D-Ala $(\rho-CDA)$ was geatly increased in an additive manner. In feeding experiments, $[1-^14C]$-D-Ala was incorporated into $\rho-CDA$at a rate of 2.2% or 2.3% of added radioactivity, indicating that exogenous D-Ala served as a precursor of the conjugate. $[1-^14C]$-L-Ala wasalso incorporated into p-CDA (0.23%) in the elicitor treated cultures. This fact suggested that at least a part of $(\rho-CDA)$ was produced from active conversion of L-Ala by the elecitation. In order to investigate a possible role of D-Ala as an elicitor of $\rpo-coumaroylamino$ acids $(\rpo-CAA)$, cold D-Ala was added together with labeled L-Ala. Although L-Ala seemed to be incorporated into $\rho-CDA$ by this treatment, the incorporation ratio was too small (0.054%) to draw a clear conclusion. However, the amount of $\rpo-coumaroylglycine$, which did not use D-Ala as a substrate, was also sightly increased by D-Ala treatment irrespective of the presence of elicitor, suggesting that exogenous D-Ala might act as an elicitor of$\rpo-CAA$ as well as a precusor substrate of $\rho-CDA$.

• 한국약용작물학회:학술대회논문집
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• 2011.09a
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• pp.408-409
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• 2011

• 한국약용작물학회:학술대회논문집
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• 2010.10a
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• pp.557-558
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• 2010