• The Plant Pathology Journal
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• 제27권4호
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• pp.310-314
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• 2011

Nonstructural protein 3 (NS3) encoded by RNA3 of Rice stripe virus (RSV), known to be a suppressor of gene silencing, was cloned and sequenced. The cloned NS3 gene is composed of 636 nucleotides encoding 211 deduced amino acids, and showed a high degree of similarity with the equivalent genes isolated from Korea, Japan and China. The NS3 gene promoted the enhancement of transient gene expression and suppressed transgene co-silencing. In the transient GFP expression via agroinfiltration, GFP expression was dramatically enhanced in terms of both protein yield and expression period in the presence of NS3. The highest accumulation of GFP protein reached to 6.8% of total soluble proteins, which corresponded to a two-fold increase compared to that obtained in the absence of NS3. In addition, NS3 significantly suppressed the initiation of GFP co-silencing induced by the additive GFP infiltration in GFP-transgenic Nicotiana benthamiana. The NS3 gene was also found to be a stronger suppressor than Cucumber mosaic virus 2b. These observations are believed to be derived from the strong suppressive effect of NS3 on gene silencing, and indicate that NS3 could be used as an effective enhancer for the rapid production of foreign proteins in plants.

• 한국패류학회지
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• 제30권4호
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• pp.391-397
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• 2014

동양달팽이의 EST 서열 4개의 클론을 어셈블리하여 추출되어진 NsCO-I (partial)서열의 코딩 영역은 852 bp, 284개의 아미노산으로 구성되어 있었다. BLAST 결과를 토대로 하여 유사도가 높은 68개의 서열을 추출 하였으며 MEGA6 프로그램을 통해 clustalW 엔진을 통해 다중서열정열을 수행하고 molecular phylogenetic analysis를 수행한 결과 Heterobranchia, Nudibranchia, Cephalaspidea, Sacoglossa, Pulmonata 등의 카테고리별로 잘 분류 되었으며 Mastigeulota kiangsinensis, Helix aspersa, Cepaea nemoralis, Elona quimperiana, Camaena cicatricosa, Cylindrus obtusus 등 육산패류들과 잘 묶인다는 사실을 확인 할 수 있었다.

• 생명과학회지
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• 제25권11호
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• pp.1304-1310
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• 2015

주기성 시계 유전자 3(period circadian clock gene 3, PER3)는 포유류에서 생물학적 주기 타이밍 시스템의 역할을 수행 한다. 이 유전자는 규칙적인 운동 체계에 의해 근육에서 전사 개시 되는 것으로 알려져 있다. 인간과 마우스에서는 본 유전자에 대해 잘 알려져 있지만, 주기 및 연주기 동안 낮의 길이에 영향을 많이 받는 말에서 운동 연관 연구는 존재하지 않는다. 운동 시 근육의 기능에 중요한 역할을 하는 PER3 유전자에 대해 대표적인 경주마인 국내산 더러브렛 품종의 운동 전과 운동 후 유전자 발현을 분석하기 위해 본 연구를 수행하였다. 그 결과, 골격근에서 PER3 유전자의 발현은 운동 전에 비해 운동 후에 유의적으로 증가하는 것으로 나타났다. 또한, 인실리코상에서 4개의 비동의성 단일 염기 변이(non-synonymous single nucleotide polymorphism, nsSNP) 분석과 이러한 nsSNP의 단백질 구조 및 기능 분석 결과, 전체 자유 에너지와 RMSD 값은 돌연변이의 원인이 될 수 있음으로 나타났다. 이 중, nsSNP–s395916798 (G72R)은 구조적 기능적 측면에서 중요한 잔기의 안정화 효과와 연관된 것을 알 수 있었다. 본 연구는 운동에 따라 더러브렛 골격근 내 PER3 발현 차이는 운동이라는 표현형에 대표될 수 있음을 확인하였다. 또한, SNP의 조합을 활용하여 운동 후 경주마의 조기 회복의 평가 지표로써 유용한 바이오마커가 될 수 있음을 시사한다.

• Journal of Microbiology
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• 제34권1호
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• pp.68-73
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• 1996

An nprX gene of Bacillus subtilis NS15-4 encoding a neutral protease was cloned and its molecular characteristics were analyzed. The complete nucleotide sequence indicated that there is an open reading frame (0RF) possibly encoding 521 amino acid polypeptide. The ORF used all codons expected two cysteine and a proline having a codon bias index (CBI) of 0.09 in Escherichia coli. There were homologous sequences to the consensus sequence of -35 and -10 regions of E. coli promoters and to a Shine-Dalgarno (SD) sequence located 25 bp downstream of a mojor transcription initiation site. Moreover, there were also five minor transcription initiation sites at 6. 7. 8. 14 and 15 nt downstream of the major site. Northern blot analysis revealed the presence of about 1.8 kb mRNA transcript in E. coli having the nprX gene. The nucleotide sequence was identified in GenBank to be a gene for a neutral protease of B. sutilis with six nucleotide difference in the ORF region. The flanking regions of the NprX ORF showed much more differences form those of other neutral protease genes except the nprE gene of B. subtilis, which has the most homology to the nprX gene, and of which the flanking regions were identical to those of the nprX gene.

• 대한바이러스학회지
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• 제27권2호
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• pp.197-207
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• 1997

To investigate the NS4 region of JEV, NS4 cDNA of K94P05 (JEV strain isolated from Korea in 1994) was amplified by RT-PCR and analyzed by sequencing PCR product. Genomic size of NS4 was 1212bp and nucleotide sequence was compared with that of other JEV strains. Nucleotide homology between JaOAr582 and K94P05 was 91.1% and that between Beijing and K94P05 was 89.8%, respectively. But the nucleotide sequence of E region of JaOAr582 and K94P05 showed 97.0% homology and that of Beijing and K94P05 did 95.8% homology. NS4 protein was expressed as a form of fusion protein by a prokaryotic expression system. The induced fusion product showed a lower molecular weight than predicted size and remained insoluble. The NS4 protein might be cleavaged by E. coli protease. Concluding above results, high hydrophobicity of the NS4 protein supported the fact that this protein played a role as a membrane component and the poor nucleotide sequence conservativity among JEV strains suggested that this region might be important to adapt each viral growth environment.

• 미생물학회지
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• 제52권2호
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• pp.166-174
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• 2016

호기적 조건에서 질산화와 탈질화를 동시에 진행하는 Alcaligenes faecalis NS13 균주를 분리하여 다양한 특성을 파악하였다. 이 균주는 $15-37^{\circ}C$ 온도에서 생장할 수 있으며 암모니움 산화율이 높고 고농도의 암모니움 환경에서도 생장이 저해되지 않고 초기 암모니움 농도 증가에 따라 제거량이 증가하였다. pH와 염분농도에 대해서도 내성 범위가 넓어 암모니움 산화가 영향을 받지 않았다. 질산화에 이어진 탈질화로 인해 질산염의 축적이 일어나지 않았으며 탈질화의 중간산물인 아산화질소는 미량 검출되었지만 배양 후 모든 질소 화합물을 측정한 결과 약 42.8%가 $N_2$로 전환된 것으로 추정되었다. 탈질화는 PCR 증폭을 통해서 탈질화에 관여하는 유전자 nitrate reductase gene, napA과 nitrous oxide reductase gene, nosZ의 존재로 뒷받침되었다. 또한 배지 내 질소의 46.4%가 NS13 균주로 동화되었기 때문에 폐수처리 시 질산화 및 탈질화 후에 슬러지로 처분한다면 실질적으로 89% 이상의 우수한 암모니움의 제거효과를 거둘 수 있을 것이다.

• 한국축산식품학회지
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• 제37권6호
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• pp.926-930
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• 2017

High-quality meat is of great economic importance to the pig industry. The 1-acylglycerol-3-phosphate-O-acyltransferase 5 (AGPAT5) enzyme converts lysophosphatidic acid to phosphatidic acid in the mitochondrial membrane. In this study, we found that the porcine AGPAT5 gene was highly expressed in muscle tissue, influencing meat characteristics, and we also identified a non-synonymous single-nucleotide polymorphism (nsSNP) (rs196952262, c.673 A>G) in the gene, associated with a change of isoleucine 225 to valine. The presence of this nsSNP was significantly associated with meat color (lightness), lower cooking loss, and lower carcass temperatures 1, 4, and 12 h after slaughter (items T1, T4, and T12 on the recognized quality scale, respectively), and tended to increase backfat thickness and the water-holding capacity. These results suggest that nsSNP (c.673A>G) of the AGPAT5 gene is a potential genetic marker of high meat quality in pigs.

• Asian Pacific Journal of Cancer Prevention
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• 제15권5호
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• pp.2291-2295
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• 2014

Although nucleolar protein nucleostemin (NS) is essential for cell proliferation and early embryogenesis and expression has been observed in some types of human cancer and stem cells, the molecular mechanisms involved in mediation of cell proliferation and cell cycling remains largely elusive. The aim of the present study was to evaluate NS as a potential target for gene therapy of human breast carcinoma by investigating NS gene expression and its effects on SKBR-3 cell proliferation and apoptosis. NS mRNA and protein were both found to be highly expressed in all detected cancer cell lines. The apoptotic rate of the pcDNA3.1-NS-Silencer group ($12.1-15.4{\pm}3.8%$) was significantly higher than those of pcDNA3.1-NS ($7.2-12.0{\pm}1.7%$) and non-transfection groups ($4.1-6.5{\pm}1.8%$, P<0.01). MTT assays showed the knockdown of NS expression reduced the proliferation rate of SKBR-3 cells significantly. Matrigel invasion and wound healing assays indicated that the number of invading cells was significantly decreased in the pcDNA3.1-NS-siRNA group (P<0.01), but there were no significant difference between non-transfected and over-expression groups (P>0.05). Moreover, RNAi-mediated NS down-regulation induced SKBR-3 cell G1 phase arrest, inhibited cell proliferation, and promoted p53 pathway-mediated cell apoptosis in SKBR-3 cells. NS might thus be an important regulator in the G2/M check point of cell cycle, blocking SKBR-3 cell progression through the G1/S phase. On the whole, these results suggest NS might be a tumor suppressor and important therapeutic target in human cancers.

• 대한임상검사과학회지
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• 제49권4호
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• pp.420-426
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• 2017

메치실린 내성 포도알균(methicillin resistant Staphylococci, MRS)는 인체 여러 부위에서 집락화 될 수 있으며 의료기관과 연관된 사람에서 흔히 분리된다. 이 연구는 밀접한 단체생활을 하는 일개 고등학교 한 개 반의 학생들 손과 그들이 사용하는 책상에서 MRS 균의 오염 정도를 평가하고자 하였다. Staphylococcus aureus가 28명의 학생의 손 중에서 2 균주가 분리되었으며 모두 메치실린에 감수성이었다. 응고인자 음성 포도알균(coagulase negative Staphylococci, CoNS)는 26 균주가(26/28, 92.9%) 분리되었으며 이들 균주 중 14 균주는 메치실린 내성균(MRCoNS)이었다. 14 MRCoNS 중 S. warneri가 가장 흔하였으며(8/14, 57%), 이들은 대부분의 $non-{\beta}-lactam$ 항생제에 감수성 이었다. 31개의 책상에서는 S. aureus는 분리되지 않았으나 26 CoNS (26/31, 83.9%)가 분리되었다. 손과 책상에서 분리된 포도알균 이외의 균은 Micrococcus와 Bacillus spp.이었다. 결론적으로 MRSA는 이번 연구에서 분리되지 않았으나 mecA 유전자를 갖고 있는 MRCoNS는 학생들의 손에서 높은 비율로 분리되었다. 손씻기와 같은 예방 교육을 강화할 뿐만 아니라 이들 균의 오염 및 보균율 등의 조사와 같은 능동적 감시 등이 이들 균의 감염예방과 전파 차단을 위해 필요할 것으로 사료된다.

• BMB Reports
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• 제37권6호
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• pp.741-748
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• 2004

The hepatitis C virus is associated with the development of liver cirrhosis and hepatocellular carcinomas. Among the 10 polyproteins produced by the virus, no function has been clearly assigned to the non-structural 5A (NS5A) protein. This study was designed to identify the hepatocellular proteins that interact with NS5A of the HCV. Yeast two-hybrid experiments were performed with a human liver cDNA prey-library, using five different NS5A derivatives as baits, the full-length NS5A (NS5A-F, amino acid (aa) 1~447) and its four different derivatives, denoted as NS5A-A (aa 1~150), -B (aa 1~300), -C (aa 300~447) and D (aa 150~447). NS5A-F, NS5A-B and NS5A-C gave two, two and 10 candidate clones, respectively, including an AHNAK-related protein, the secreted frizzled-related protein 4 (SFRP4), the N-myc downstream regulated gene 1 (NDRG1), the cellular retinoic acid binding protein 1 (CRABP-1), ferritin heavy chain (FTH1), translokin, tumor-associated calcium signal transducer 2 (TACSTD2), phosphatidylinositol 4-kinase (PI4K) and $centaurin{\delta}$ 2 ($CENT{\delta}2$). However, NS5A-A produced no candidates and NS5A-D was not suitable as bait due to transcriptional activity. Based on an in vitro binding assay, CRABP-1, PI4K, $CENT{\delta}2$ and two unknown fusion proteins with maltose binding protein (MBP), were confirmed to interact with the glutathione S-transferase (GST)/NS5A fusion protein. Furthermore, the interactions of CRABP-1, PI4K and $CENT{\delta}2$ were not related to the PXXP motif (class II), as judged by a domain analysis. While their biological relevance is under investigation, the results contribute to a better understanding of the possible role of NS5A in hepatocellular signaling pathways.