• Journal of Ginseng Research
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• 제42권4호
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• pp.496-503
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• 2018

Background: Korean ginseng (Panax ginseng) plays an anti-inflammatory role in a variety of inflammatory diseases such as gastritis, hepatitis, and colitis. However, inflammation-regulatory activity of the calyx of the P. ginseng berry has not been thoroughly evaluated. To understand whether the calyx portion of the P. ginseng berry is able to ameliorate inflammatory processes, an ethanolic extract of P. ginseng berry calyx (Pg-C-EE) was prepared, and lipopolysaccharide-activated macrophages and HEK293 cells transfected with inflammation-regulatory proteins were used to test the anti-inflammatory action of Pg-C-EE. Methods: The ginsenoside contents of Pg-C-EE were analyzed by HPLC. Suppressive activity of Pg-C-EE on NO production, inflammatory gene expression, transcriptional activation, and inflammation signaling events were examined using the Griess assay, reverse transcription-polymerization chain reaction, luciferase activity reporter gene assay, and immunoblotting analysis. Results: Pg-C-EE reduced NO production and diminished mRNA expression of inflammatory genes such as cyclooxygenase-2, inducible NO synthase, and tumor necrosis factor-${\alpha}$ in a dose-dependent manner. This extract suppressed luciferase activity induced only by nuclear factor-${\kappa}B$. Interestingly, immunoblotting analysis results demonstrated that Pg-C-EE reduced the activities of protein kinase B (AKT)1 and AKT2. Conclusion: These results suggest that Pg-C-EE may have nuclear-factor-${\kappa}B$-targeted anti-inflammatory properties through suppression of AKT. The calyx of the P. ginseng berry is an underused part of the ginseng plant, and development of calyx-derived extracts may be useful for treatment of inflammatory diseases.

• 미생물학회지
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• 제28권1호
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• pp.19-26
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• 1990

To examine the effects of sequence variations near the transcriptional start site on the rate of formation of the open complexes at bacteriophage $\lambda P_{R}$ promoter, two mutant promoters were created by site-specific mutagenesis using synthetic oligonucleotides. Mutant I coatains changes at positions -3 and -4 from TT to CC, thus having a 6-bp long G/C stretch between -10 region and transciptional start site (+1). Mutant II has changes at positions -5 and -6 from GG to AA, thereby having a 9-bp long A/T stretch between positions -11 and -3. Selective filter binding assays were performed to measure the rate of formation of the open complexes between the wild-type or two mutant $P_{R}$ promoters on 664 bp fragments and E. coli RNA polymerase at two temperatures. At 37.deg.C, the wild-type and two mutants showed similar rates for the formation of open complex. The second order rate constant $k_{a}$ and $\tau _{int}$, as determined from the .tau.-plot analysis, were $(6.0\pm0.4)\times10^{6}M^{-1}sec^{-1}$ and $11\pm5$sec, respectively. At 18.deg.C, however, the wild-type and two mutant promoters showed differences in the kinetic parameters. k for the wild-type promoter was (2.2$\pm$0.1)\times 10^{6}M^{-1}sec^{-1}$ and $\tau _{int}$ was 76$\pm$sec. Mutant I and II exhibited differences mainly in the rate of isomerization ($\tau_{int,I}=91\pm$10 sec, int,II=34$\pm$ sec), whereas the second order rate constant $k_{a}$ was similar to the wild type value. This result implies that at $18^{\circ}C$, the isomerization rate is determined by both protein conformational change and DNA melting, which are separable kinetically according to the 3-step mechanism of Roe et al.(1984,1985), and that the base changes affected mainly the rate of DNA melting as predicted.lting as predicted.

• BMB Reports
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• 제33권6호
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• pp.519-524
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• 2000

NF1 proteins are a family of DNA binding proteins which consist of two separate domains, N-terminal DNA binding domain and C-terminal transcription activation domain. The N-terminal 220 amino acids are highly conserved and are also known to mediate dimerization of NF1 proteins. The C-terminal regions of different type of NF1 proteins are heterogeneous and responsible for transcriptional activation. In this study, we tested the interaction between different domains of rat NF1-A protein by yeast two hybrid analysis and observed the interaction between C-terminal regions of NF1-A which do not contain the N-terminal dimerization domain. Our results showed that the C-terminal region of rat NF1-A between residues 231 and 509 strongly interacted not only with itself, but also with human NF1/CTF1 which is a different type of NF1. When the C-terminal region was divided into two fragments, one from residue 231 to 447 and the other from 448 to 509, the two fragments were able to interact with the C-terminal region of NF1-A significantly. This indicates that both fragments contain independent interaction domains. Analysis of the interactions with alanine substituted fragments showed that substitutions of the heptasequence, SPTSPTY of NF1-A, affected interaction between NF1 proteins. Our results strongly suggest that C-terminal regions may also be important for the formation of homo- and heterodimers in addition to the N-terminal dimerization domain. Also, the heptasequence motif may play some roles in dimer formation.

• 한국버섯학회지
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• 제9권3호
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• pp.91-95
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• 2011

느타리버섯은 가장 중요한 식용버섯 중 하나이다. 느타리버섯은 Pseudomonas tolaasii에 의한 세균성 갈변병에 매우 감수성이므로, 저항성 품종을 만들기 위한 노력의 하나로 누에에서 분리된 항 세균성 단백질인 누에신을 느타리버섯에서 과발현시키고자 하였다. 누에신 cDNA는 여름 느타리버섯의 ${\beta}$-tubulin 프로모터에 결합되어 pTRura3-2 vector와 함께 우라실 영양요구성 돌연변이 균주에 형질전환되었다. 누에신 cDNA가 형질전환된 느타리버섯을 genomic PCR과 Southern blot을 통하여 분리할 수가 있었으며, 이들 중 3개의 형질전환체가 누에신 유전자를 발현시킴을 확인하였다. 그러나 이들 형질전환체들에서 누에신 단백질을 검출할 수 없었으며, 또한 항 세균 효과도 확인할 수 없었다. 이들 결과는 형질전환기술을 이용한 병 저항성 개발 가능성을 보여주고 있다.

• 미생물학회지
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• 제53권1호
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• pp.49-54
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• 2017

Sus1 / ENY2는 진핵생물에 잘 보존된 작은 단백질로 염색사 리모델링과 mRNA 생성과정에 관여한다. Sus1 단백질은 전사 보조활성자인 SAGA 복합체와 핵공과 연관된 TREX2 복합체 등, 핵에 존재하는 2개 복합체의 구성 요소이다. 분열효모 Schizosaccharomyces pombe의 유전체 데이터에서 Sus1의 이종상동체를 찾아 기능을 분석하였다. 4분체 분석 결과 이 유전자는 생장에 필수적이 아니었으나, Sus1 유전자를 결실시키면 낮은 온도에서 생장이 느렸으며, $poly(A)^+$ RNA도 핵 안에 약간 축적되는 표현형을 보였다. 또한 Sus1-GFP 단백질은 주로 핵에 존재하였다. Yeast two-hybrid 분석과 공동면역침전 실험에서 Sus1 단백질은 TREX-2 복합체의 또 다른 구성인자인 Sac3와 상호작용을 하였다. 이와 같은 결과들은 S. pombe의 Sus1 단백질도 역시 TREX-2 복합체의 구성인자로 mRNA 방출에 관여하고 있음을 시사한다.

• Journal of Microbiology and Biotechnology
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• 제13권4호
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• pp.598-606
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• 2003

Salmonella encodes an enterotoxin (Stn) which possesses biological activity similar to the cholera toxin. Stn contributes significantly to the overall virulence of S. typhimurium in a murine model. The production of Stn is enhanced in a high-osmolarity medium and by contact with epithelial cells. In the present study, the in vitro and in vivo transcriptional regulations of the sin promoter revealed two promoters, P1 and P2. The P1 promoter identified by a primer extension analysis of stn mRNA exhibited a switching mechanism in vivo. Depending on the growth stage, transcription was initiated from different start sites termed $P1_S\;and\;P1_E$. $P1_S$, recognized by RNA polymerase containing ${\sigma}^S(E{\sigma}^S),\;and\;P1_E$, recognized by $E{\sigma}^70$, were activated during the stationary and exponential phases, respectively, while $P1_S\;and\;P1_E$ were both negatively regulated by CRPㆍcAMP and H-NS. Results revealed that $P1_S$ was the responsible promoter activated under a high osmolarity and low pH. The P2 promoter was identified 45 nucleotides downstream from $P1_E$ and negatively controlled by CRPㆍcAMP in vitro. No P2 activity was detected in vivo. The regulation of stn expression monitored using a Pstn::egfp fusion indicated that $E{\sigma}^S$ was required for the induction of stn and various factors were involved in stn regulation inside animal cells.

• 미생물학회지
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• 제54권3호
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• pp.175-187
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• 2018

점액세균은 포식활동, 자기방어, 세포 간 신호전달 및 아직까지 알려지지 않은 다른 기능을 위해 다양한 이차대사산물을 생산한다. 점액세균에서 분리된 많은 이차대사산물들은 독특한 작용기작을 가지며 항암, 항세균, 항진균 등과 같은 약학적으로 유용한 생리활성을 보인다. 따라서 전 세계적으로 많은 점액세균 균주들이 분리되었고 이들로부터 다양한 생리활성물질들이 탐색되었다. 하지만 16S rRNA 데이터베이스 분석에 의하면 야생에는 지금까지 분리된 종류 이외에도 다양한 점액세균 종류들이 존재할 것으로 추정되며, 유전체 서열 분석에 의하면 각 점액세균들은 기존에 알려진 물질보다 더 많은 물질을 생산할 수 있는 능력이 있는 것으로 나타났다. 본 총설에서는 점액세균 유래 이차대사산물들과 이들의 유전자, 점액세균에서의 기능, 생합성 유전자의 발현을 조절하는 전사조절인자 등에 대한 최근까지의 연구 현황을 살펴보았다.

• 식물조직배양학회지
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• 제24권3호
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• pp.161-165
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• 1997

옥수수의 색소합성을 조절하는 pattern allele의 하나인 R-mb유전자의 구조와 유전적 분석을 수행하였다. R-mb 유전자의 유전분석을 수행한 결과를 검토하여 볼 때 후대로 진행됨에 따라 색소 발현빈도_의 감소경향을 보이고 있었다. 또한 R-mb 유전자가 몇 개의 R subcomplex로 존재하는가를 알기 위해서는 우선 R specific probe인 pR-nj:1를 이용하여 Southern blot hybridization을 실시한 결과 약 3.9kb 및 약 7.75kb영역에서 2개의 band가 관찰되었다. R-mb 유전자를 클로닝하기 위하여 λFIXIIvector를 이용하여 library를 만들고 이로부터 mb-II, III, V, Ⅵ, Ⅶ 등 5개의 clone을 3차의 screen을 거쳐 확보하고 이중 mb-II 및 mb-Ⅵ를 중심으로 제한효소지도를 작성하였으며, 이 유전자의 구조와 기타 R locus관련 유전자들과 비교하였으며, 이러한 두 개의 R 요소가 어떻게 색소발현에 영향을 미치는가에 대에 검토하였다.

• 방사선산업학회지
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• 제4권4호
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• pp.325-334
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• 2010

Transcriptional regulation in response to salt in mutant lines was investigated using oligonucleotide microarrays. In order to characterize the salt-responsive genes in rice, the expression profiles of transcripts that responded to salt-treatment were monitored using the microarrays. In the microarray analysis, among 37,299 reliable genes, 5,101, 2,758 and 2,277 genes were up-regulated by more than 2-fold using the salt treatment, while the numbers of down-regulated genes were 4,619, 3,234, and 1,878 in the WT, ST-495, and ST-532, respectively. From genotype changes induced by gamma ray mutagenesis, 3,345 and 2,397 genes were up-regulated, while 2,745 and 2,075 genes were down-regulated more than 2-fold in the two untreated mutants lines compared with untreated WT, respectively. A total of 3,108 and 2,731 genes were up-regulated more than 2-fold, while 3,987 and 3,660 genes were down-regulated by more than 2-fold in the salt treatment of the two mutants lines compared with the salt treated WT, respectively. The expressions of membrane transporter genes such as OsAKT1, OsKUP, and OsNAC increased more severely in ST-495 and ST-532 than in the WT. The expressions of the proline accumulation related genes such as OsP5CS and OsP5CR were also markedly increased in the salt tolerant mutants when compared to the WT plant.

• Journal of Periodontal and Implant Science
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• 제51권6호
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• pp.386-397
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• 2021

Purpose: MicroRNAs (miRNAs) are epigenetic post-transcriptional regulators that modulate gene expression and have been identified as biomarkers for several diseases, including cancer. This study aimed to systematically review the relationship between miRNAs and periodontal disease in humans, and to evaluate the potential of miRNAs as diagnostic and prognostic biomarkers of disease. Methods: The review was conducted following the Preferred Reporting Items for Systematic Reviews and Meta-Analysis guidelines (reference number CRD42020180683). The MEDLINE, Scopus, Cochrane Library, Embase, Web of Science, and SciELO databases were searched for clinical studies conducted in humans investigating periodontal diseases and miRNAs. Expression levels of miRNAs across the different groups were analysed using the collected data. Results: A total of 1,299 references were identified in the initial literature search, and 23 articles were finally included in the review. The study designs were heterogeneous, which prevented a meta-analysis of the data. Most of the studies compared miRNA expression levels between patients with periodontitis and healthy controls. The most widely researched miRNA in periodontal diseases was miR-146a. Most studies reported higher expression levels of miR-146a in patients with periodontitis than in healthy controls. In addition, many studies also focused on identifying target genes of the differentially expressed miRNAs that were significantly related to periodontal inflammation. Conclusions: The results of the studies that we analysed are promising, but diagnostic tests are needed to confirm the use of miRNAs as biomarkers to monitor and aid in the early diagnosis of periodontitis in clinical practice.