• Journal of Microbiology
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• 제44권4호
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• pp.432-438
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• 2006

Twelve actinomycete strains were isolated from Egyptian soil. The isolated actinomycete strains were then screened with regard to their potential to generate antibiotics. The most potent of the producer strains was selected and identified. The cultural and physiological characteristics of the strain identified. the strain as a member of the genus Streptomyces. The nucleotide sequence of the 16S rRNA gene (1.5kb) of the most potent strain evidenced a 99% similarity with Streptomyces spp. and S. aureofaciens 16S rRNA genes, and the isolated strain was ultimately identified as Streptomyces sp. MAR01. The extraction of the fermentation broth of this strain resulted in the isolation of one major compound, which was active in vitro against gram-positive, gram-negative representatives and Candida albicans. The chemical structure of this bioactive compound was elucidated based on the spectroscopic data obtained from the application of MS, IR, UV, $^1H$ NMR, $^{13}C$ NMR, and elemental analysis techniques. Via comparison to the reference data in the relevant literature and in the database search, this antibiotic, which had a molecular formula of $C_{19}H_{29}NO_2$ and a molecular weight of 303.44, was determined to differ from those produced by this genus as well as the available known antibiotics. Therefore, this antibiotic was designated Meroparamycin.

• Animal Bioscience
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• 제36권6호
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• pp.851-860
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• 2023

Objective: This study aims to evaluate the target genes of gga-miR-20a-5p and the regulated immune responses in the chicken macrophage cell line, HD11, by the exosome-mediated delivery of miR-20a-5p. Methods: Exosomes were purified from the chicken macrophage cell line HD11. Then, mimic gga-miR-20p or negative control miRNA were internalized into HD11 exosomes. HD11 cells were transfected with gga-miR-20a-5p or negative control miRNA containing exosomes. After 44 h of transfection, cells were incubated with or without 5 ㎍/mL poly(I:C) for 4 h. Then, expression of target genes and cytokines was evaluated by quantitative realtime polymerase chain reaction. Results: Using a luciferase reporter assay, we identified that gga-miR-20a-5p directly targeted interferon gamma receptor 2 (IFNGR2), mitogen-activated protein kinase 1 (MAPK1), mitogen-activated protein kinase kinase kinase 5 (MAP3K5), and mitogen-activated protein kinase kinase kinase 14 (MAP3K14). Moreover, the exosome-mediated delivery of gga-miR-20a-5p successfully repressed the expression of IFNGR2, MAPK1, MAP3K5, and MAP3K14 in HD11 cells. The expressions of interferon-stimulated genes (MX dynamin like GTPase 1 [MX1], eukaryotic translation initiation factor 2A [EIF2A], and oligoadenylate synthase-like [OASL]) and proinflammatory cytokines (interferon-gamma [IFNG], interleukin-1 beta [IL1B], and tumor necrosis factor-alpha [TNFA]) were also downregulated by exosomal miR-20a-5p. In addition, the proliferation of HD11 cells was increased by exosomal miR-20a-5p. Conclusion: The exosome-mediated delivery of gga-miR-20a-5p regulated immune responses by controlling the MAPK and apoptotic signaling pathways. Furthermore, we expected that exosomal miR-20a-5p could maintain immune homeostasis against highly pathogenic avian influenza virus H5N1 infection by regulating the expression of proinflammatory cytokines and cell death.

• Molecules and Cells
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• 제41권5호
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• pp.390-400
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• 2018

Studies have revealed that miR-103a-3p contributes to tumor growth in several human cancers, and high miR-103a-3p expression is associated with poor prognosis in advanced gastric cancer (GC) patients. Moreover, bioinformatics analysis has shown that miR-103a-3p is upregulated in The Cancer Genome Atlas (TCGA) stomach cancer cohort. These results suggest that miR-103a-3p may function as an oncogene in GC. The present study aimed to investigate the role of miR-103a-3p in human GC. miR-103a-3p expression levels were increased in 33 clinical GC specimens compared with adjacent nontumor stomach tissues. Gain- and loss-of-function studies were performed to identify the correlation between miR-103a-3p and tumorigenesis in human GC. Inhibiting miR-103a-3p suppressed GC cell proliferation and blocked the S-G2/M transition in MKN-45/SGC-7901 cells, whereas miR-103a-3p overexpression improved GC cell proliferation and promoted the S-G2/M transition in vitro. Bioinformatics and dual-luciferase reporter assays confirmed that ATF7 is a direct target of miR-103a-3p. Analysis of the TCGA stomach cancer cohort further revealed that miR-103a-3p expression was inversely correlated with ATF7 expression. Notably, silencing ATF7 showed similar cellular and molecular effects as miR-103a-3p overexpression, namely, increased GC cell proliferation, improved CDK2 expression and decreased P27 expression. ATF7 overexpression eliminated the effects of miR-103a-3p expression. These findings indicate that miR-103a-3p promotes the proliferation of GC cell by targeting and suppressing ATF7 in vitro.

• 한국응용곤충학회지
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• 제39권1호
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• pp.5-12
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• 2000

임의증폭 다형 DNA(RAPD)와 미토콘드리아 12S, 16S rRNA 유전자의 제한단편 DNA 표식자에 의해 한국에서 발생하는 담배가루이 개체군들의 biotype을 판별하였다. 진천의 장미 온실과 서울 내곡동의 포인세티아 온실에서 발생한 담배가루이는 일본, 이스라엘, 호주의 B biotype 과 동일한 DNA 단편들을 보유하였다. 여러 지역의 노지 콩 (Glycine max), 고구마 (Ipomea batatas), 들깨 (Perilla frutescens)에서 채집된 담배가루이 개체군들은 일본 시코쿠의 인동덩굴(Lonicera japonica)에서 채집된 담배가루이와 같은 DNA로 표식되었다. 이들 non-B biotype은 한국, 일본 등 극동아시아 지역에 고유한 계통으로 보인다. 최근 한국에서 발견된 담배가루이 Bemisia tabaci(Gennadius)의 주사전자현미경 관찰에 의한 형태적 특정을 온실 원예작물의 주요해충인 온실가루이 Trialeurodes vaporariorum (Westwood)와 비교하여 기재하였다.

• Animal cells and systems
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• 제3권3호
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• pp.295-301
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• 1999

The nucleotide sequence of a 447 base pair fragment in the mitochondrial cytochrome b gene and the complete sequence of the mitochondrial 12S ribosomal RNA gene, 938 bp, were analyzed to infer inter- and intraspecific genetic relationships of Hyla japonica and H. suweonensis from Korea and H, japonica from Japan. In the mitochondrial cytochrome b gene, genetic differentiation among H. japonica populations were 9.62% and 15.66% between H. japonica and H. suweonensis. Based on the Tamura-Nei distance, the level of sequence divergence ranged from 0.45% to 2.75% within Korean H. japonica, while 8.31%-8.87% between Korean and Japanese H. japonica and 11.51%-12.46% between H. japonica and H. suweonensis. In the neigh-bor-joining tree, Korean populations of H. japonica were clustered first at 2.22% and followed by Japanese H. japonica and H. suweonensis at 8.51% and 12.29%, respectively. In mitochondrial 12S rRNA gene, genetic differentiation between H. japonica and H. suweonensis nras 7.17% (68 bp) including 7 gaps. Based on Tamura-Nei distance, the level of sequence divergence ranged 3.53% between Korean and Japanese H. japonica and from 4.93% to 5.41% between H. japonica and H. suweonensis. Phenogram pattern of the 12S rRNA gene sequence corresponded with that of the mitochondrial cytochrome b gene.

• Fisheries and Aquatic Sciences
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• 제16권3호
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• pp.165-169
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• 2013

We report on mycobacteriosis in an imported tropical ornamental fish Macropodus opercularis commonly known as the paradise fish. Mass mortality occurred in paradise fish imported to Korea from Southeast Asia in 2008. The affected fish did not show any outward clinical signs, but enlargement of the spleen, kidneys, and liver was observed on dissection. Histopathological examination revealed numerous granulomas in the spleen, and acid-fast bacilli were observed in the centers of the granulomas. About 65% of spleen DNA samples were PCR positive using mycobacteria-specific primers targeting the 16S rRNA and hsp65 genes. The nucleotide identities of the 16S rRNA and hsp65 genes with those of Mycobacterium marinum were 99.5% and 99.4%, respectively. Although the bacterium was not cultured, the molecular diagnosis and histopathological findings were consistent with mycobacteriosis in paradise fish.

• 한국미생물·생명공학회지
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• 제48권2호
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• pp.193-204
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• 2020

Sixteen acetic acid bacteria (AAB) were isolated from blueberries and citric fruits of the Salto Grande region (Concordia, Entre Rios, Argentina) using enrichment techniques and plate isolation. Enrichment broths containing ethanol and acetic acid enabled maximum AAB recovery, since these components promote their growth. Biochemical tests allowed classification of the bacteria at genus level. PCR-RFLP of the 16S rRNA and PCR-RFLP of the 16S-23S rRNA intergenic spacer allowed further classification at the species level; this required treatment of the amplified products of 16S and 16S-23S ITS ribosomal genes with the following restriction enzymes: AluI, RsaI, HaeIII, MspI, TaqI, CfoI, and Tru9I. C7, C8, A80, A160, and A180 isolates were identified as Gluconobacter frateurii; C1, C2, C3, C4, C5, C6, A70, and A210 isolates as Acetobacter pasteurianus; A50 and A140 isolates as Acetobacter tropicalis; and C9 isolate as Acetobacter syzygii. The bacteria identified by 16S rRNA PCR-RFLP were validated by 16S-23S PCR-RFLP; however, the C1 isolate showed different restriction patterns during identification and validation. Partial sequencing of the 16S gene resolved the discrepancy.

• International Journal of Oral Biology
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• 제37권3호
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• pp.146-152
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• 2012

MicroRNAs (miRNAs, miRs) are about 21-25 nucleotides in length and regulate mRNA translation by base pairing to partially complementary sites, predominantly in the 3'-untranslated region (3'-UTR) of the target mRNA. In this study, the expression profile of miRNAs was compared and analyzed for the establishment of miRNA-related odontoblast differentiation using MDPC-23 cells derived from mouse dental papilla cells. To determine the expression profile of miRNAs during the differentiation of MDPC-23 cells, we employed miRNA microarray analysis, quantitative real-time PCR (qRT-PCR) and Alizaline red-S staining. In the miRNA microarray analysis, 11 miRNAs were found to be up- or down-regulated more than 3-fold between day 0 (control) and day 5 of MDPC-23 cell differentiation among the 1,769 miRNAs examined. In qRT-PCR analysis, the expression levels of two of these molecules, miR-194 and miR-126, were increased and decreased in the control MDPC-23 cells compared with the MDPC-23 cells at day 5 of differentiation, respectively. Importantly, the overexpression of miR-194 significantly accelerated mineralization compared with the control cultures during the differentiation of MDPC-23 cells. These results suggest that the miR-194 augments MDPC-23 cell differentiation, and potently accelerates the mineralization process. Moreover, these in vitro results show that different miRNAs are deregulated during the differentiation of MDPC-23 cells, suggesting the involvement of these genes in the differentiation and mineralization of odontoblasts.

• 생명과학회지
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• 제22권1호
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• pp.110-116
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• 2012

누룩은 탁주와 약주의 제조를 위한 당화효소와 알코올발효를 위한 미생물의 공급원으로서 제품의 맛과 품질을 결정하는 중요한 역할을 한다. 본 연구에서는 산성누룩의 세균과 진균의 다양성을 조사하기 위해 순수분리 종과 16S 및 28S rRNA gene를 대상으로 한 PCR-DGGE를 이용한 분석을 수행하였다. 누룩 내 세균의 수는 $2.7{\times}10^9$ CFU/g이었으며 순수분리와 PCR-DGGE 분석에서 우점종은 Kocuria spp., Pantoea spp., Lactobacillus spp., Pediococcous spp., Weissella spp., Staphylococcus spp. 그 외 endophytic bacterium, uncultured gamma-proteobacteria, uncultured Cyanobacteria와 Actinobacteria였다. PCR-DGGE profile에서 주된 우점종은 Pediococcous pentosaceus와 uncultured Cyanobacteria 이었다. 누룩 내 진균의 수는 $3.5{\times}10^8$ CFU/g이었으며 순수분리와 PCR-DGGE 분석에서 우점종은 Trichomonascus spp., Pichia spp., Torulaspora spp., Wickerhamomyces spp., Sacharomycopsis spp., Lichtheimia spp., Mucor spp., Rhizopus spp., Aspergillus spp., Cladosporium spp.였다. PCR-DGGE profile에서 주된 우점종은 Pichia kudriavzevii와 Aspergillus oryzae이었다. PCR-DGGE 기술은 본 연구에서 누룩의 미생물군집을 평가하기 위해 처음으로 사용되었으며 미생물 다양성을 설명하는 데 효과적임을 입증하였다.

• 한국자원식물학회지
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• 제19권1호
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• pp.161-168
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• 2006

제주도에 자생하는 부채 선인장인 백년초의 기원 규명을 목적으로 ITS primer를 이용하여 685 bp의 ITS 영역을 분리하였다. ITS 영역의 염기서열을 분석한 결과 18S rRNA의 길이는 54 bp, 26S rRNA는 55 bp, ITS1은 193 bp, ITS2는 220 bp로 구성되어 있었다. 백년초 ITS 영역은 기존에 보고된 Cucurbitoideae 식물들의 ITS 영역에 비하여 ITS2 스페이서 영역의 239-254 bp보다는 다소 짧았다. 그러나 이들 스페이서 영역의 GC 함량은 백년초의 경우 ITS1은 66.8%, ITS2의 경우에는 67.7%로 Cucurbitoideae 식물들에서 보다 높은 GC 함량을 나타내었다. 백년초 선인장의 rDNA 영역에 가장 높은 상동성을 나타낸 것은 같은 Opuntioideae에 속하는 Pereskiopsis porteri(L78037)로 95%의 유사도를 나타내었다. 백년초 rDNA Clustal W 프로그램을 이용하여 유연관계를 조사한 결과 같은 Opuntioideae에 속하는 Pereskiopsis porteri(L78037)와 같은 cluster로 분리되었다.