• Title/Summary/Keyword: 6S RNA

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Morphogenetic Identification of Eel's Larva (Leptocephalus) Collected by Set net in Namhae, Korea (남해 정치망에서 채집한 엽상자어(Leptocephalus)의 형태 및 유전학적 특성)

  • Chang-Gi Hong;Kyeong-Ho Han
    • Journal of Marine Life Science
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    • v.8 no.2
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    • pp.128-135
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    • 2023
  • The present study was tried to identify whether the eel's larva was close to a conger (Conger myriaster), a pipe conger (Muraenesox cinereus) or four species of Anguilla. Experimental fishes were collected by set net in the gulf of enggang, Namhae, Korea from May to June. Their morphological characteristics were compared with adult fishes of a conger, a pipe conger and four species of Anguilla. For genetic classification, DNA was isolated and amplified by using 12S rRNA and 16S rRNA primer set. The PCR products were direct sequencing in both directions. The nucleotide sequences were analyzed using softwares. As results of morphological measurement on eel's larva, the percentages of head length and preanal length against total length were similar with a conger. Based on the nucleotide sequences, the phylogenetic tree also revealed a close relationship to a conger. Therefore, eel's larva, caught in Namhae from May to June, was identified into a conger's larva.

Bacterial Community Analysis of Lake Soyang in Winter by Using 16S and 23S rRNA-targeted Probes (16S와 23S rRNA에 결합하는 probe를 이용한 겨울철 소양호 세균 군집 구조의 분석)

  • Hong, Sun-Hee;Byeon, Myeong-Seop;Ahn, Tae-Seok
    • Korean Journal of Microbiology
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    • v.33 no.4
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    • pp.257-261
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    • 1997
  • To scrutinize the bacterial community composition of Lake Soyang in winter, bacterial numbers belonging to Eubacteria, Proteobacteria and Cytophaga-Flavobacterium group were estimated by using 16S and 23S rRNA targeted oligonucleotide probes. Total bacterial numbers ranged from $0.7{\times}10^6$ to $1.1{\times}10^6cells{\cdot}ml^{-1}$, and vertical profile of total bacteria showed a peak at 5 m depth. The ratio of eubacteria to total bacteria were 34~90% and at 5 m and 10 m depths those were low exhibiting, 39 and 34%, respectively. The percentage of proteobacteria ${\alpha}$-group ranged 10.8~28.7%, ${\beta}$-group 4.5~53.5%, ${\gamma}$-group 4.9~35.5% and Cytophaga-Flavobacterium group 6.1~21.1%. The dominant groups were ${\beta}$-group at 0, 2 and 5 m, ${\gamma}$-group at 10 m, ${\alpha}$-group at 30 m and Cytophaga-Flavobacterium group at 50 m depth. In winter season, Lake Soyang can be divided into three layer, 0~2 m, 5~10 m and 30~50 m, by the bacteria community composition. By this method, new informations about aquatic ecosystem were developed.

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Identification of Bacteria by Sequence Analysis of 16S rRNA in Testes of Jeju Horses (제주마 고환내 세균의 16S rRNA 염기서열 분석을 이용한 동정)

  • Park, Yong-Sang;Kim, Nam-Young;Han, Sang-Hyun;Park, Nam-Geon;Ko, Moon-Suck;Cho, Won-Mo;Chae, Hyun-Seok;Cho, In-Chul;Cho, Sang-Rae;Woo, Jae-Hoon;Kang, Tae-Young
    • Journal of Veterinary Clinics
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    • v.31 no.1
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    • pp.36-39
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    • 2014
  • Many bacteria colonized in the horse semen affect quality of the sperm and some may cause infection in the mare reproductive tract and infertility of susceptible mare. This study was initiated to determine the prevalence of bacteria in testes of Jeju horses by determining rRNA sequence. The samples were swabed from the testes of nine Jeju horses (aged from 8 to 12 months after birth). Bacteria isolated from testes were identified by 16S rDNA sequencing. 1.6-kbp PCR products for 16S rRNA coding region were obtained using the universal primers. The PCR products were further purified and sequenced. Maximum similar species were found by BLAST search in the GenBank DNA database. BLAST results showed that the sequences were similar to those of Acinetobacter sp (A. schindleri, A. ursingii)., Bacillus cereus, Corynebacterium glutamicum, Escherichia coli, Gamma proteobacterium, Micrococcus luteus, Pseudomonas mendocina, Shigella sonnei, Sphingomonas sp., Staphylococcus sp (S. cohnii, S. saprophyticus, S. xylosus)., and Stenotrophomonas maltophilia. DNA sequences for 16S rRNA is provided useful informations for species identification of pathogenic microorganisms for the reproductive organs in horses.

Effects of Sasa quelpaertensis Extract on mRNA and microRNA Profiles of SNU-16 Human Gastric Cancer Cells (SNU-16 위암 세포의 mRNA 및 miRNA 프로파일에 미치는 제주조릿대 추출물의 영향)

  • Jang, Mi Gyeong;Ko, Hee Chul;Kim, Se-Jae
    • Journal of Life Science
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    • v.30 no.6
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    • pp.501-512
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    • 2020
  • Sasa quelpaertensis Nakai leaf has been used as a folk medicine for the treatment of gastric ulcer, dipsosis, and hematemesis based on its anti-inflammatory, antipyretic, and diuretic characteristics. We have previously reported the procedure for deriving a phytochemical-rich extract (PRE) from S. quelpaertensis and how PRE and its ethyl acetate fraction (EPRE) exhibits an anticancer effect by inducing apoptosis in various gastric cancer cells. To explore the molecular targets involved in this apoptosis, we investigated the mRNA and microRNA profiles of EPRE-treated SNU-16 human gastric cancer cells. In total, 2,875 differentially expressed genes were identified by RNA sequencing, and gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses indicated that the EPRE-modulated genes are associated with apoptosis, mitogen-activated protein kinase, inflammatory response, tumor necrosis factor signaling, and cancer pathways. Subsequently, protein-protein interaction network analysis confirmed interactions among genes associated with cell death and apoptosis, and 27 differentially expressed microRNAs were identified by further sequencing. Here, GO and KEGG pathway analysis revealed that EPRE modified the expression of microRNAs associated with the cell cycle and cell death, as well as signaling of tropomyosin-receptor-kinase receptor, transforming growth factor-b, nuclear factor kB, and cancer pathways. Taken together, these results provide insight into the mechanisms underlying the anticancer effect of EPRE.

Effects of Different Types of Dietary Fat on Muscle Atrophy According to Muscle Fiber Types and PPAR${\delta}$ Expression in Hindlimb-Immobilized Rats (지방의 종류가 다른 식이의 섭취가 하지고정 흰 쥐의 근 섬유별 근 위축과 PPAR${\delta}$ 활성에 미치는 영향)

  • Lee, Ho-Uk;Park, Mi-Na;Lee, Yeon-Sook
    • Journal of Nutrition and Health
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    • v.44 no.5
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    • pp.355-365
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    • 2011
  • This study investigated how dietary fat affects muscle atrophy and lipid metabolism in various muscles during hindlimb immobilization in rats. Twenty-four male Sprague?Dawley rats had their left hindlimb immobilized and were divided into four groups by dietary fat content and composition. The contralateral hindlimb (control) was compared with the immobilized limb in all dietary groups. Rats (n = 6/group) were fed a 4% corn oil diet (CO), 2.6% corn oil + 1.4% fish oil diet (FO), 30% corn oil diet (HCO), or a 30% beef tallow diet (HBT)after their hind limbs were immobilized for 10 days. Data were collected for the gastrocnemius, plantaris and soleus muscles. Muscle atrophy was induced significantly after 10 days of hindlimb immobilization, resulting in significantly decreased muscle mass and total muscle protein content. The protein levels of peroxisome proliferator activated receptor ${\delta}$ (PPAR${\delta}$) in the plantaris, gastrocnemius, and soleus increased following hindlimb immobilization irrespective of dietary fat intake. Interestingly, the PPAR${\delta}$ mRNA level in the plantaris decreased significantly in all groups and that in the FO group was lower than that in the other groups. The soleus PPAR${\delta}$ mRNA level decreased significantly following hindlimb immobilization in the FO group only. Muscle carnitine palmitoyl transferase 1 (mCPT1) mRNA level was not affected by hindlimb immobilization. However, the mCPT1 mRNA level in the FO group was significantly lower in the plantaris but higher in the soleus than that in the other groups. The pyruvate dehydrogenase kinase 4 (PDK4) mRNA level in the plantaris decreased significantly, whereas that in the soleus increased significantly following hindlimb immobilization. The plantaris, but not soleus, PDK4 mRNA level was significantly higher in the FO group than that in the CO group. The increased PPAR${\delta}$ protein level following hindlimb immobilization may have suppressed triglyceride accumulation in muscles and different types of dietary fat may have differentially affected muscle atrophy according to muscle type. Our results suggest that ${\omega}$-3 polyunsaturated fatty acids may suppress muscle atrophy and lipid accumulation by positively affecting the expression level and activity of PPAR${\delta}$ and PPAR${\delta}$-related enzymes, which are supposed to play an important role in muscle lipid metabolism.

Integrin-linked Kinase Functions as a Tumor Promoter in Bladder Transitional Cell Carcinoma

  • Wang, De-Lin;Lan, Jian-Hua;Chen, Liang;Huang, Biao;Li, Zeng;Zhao, Xiu-Min;Ma, Qiang;Sheng, Xia;Li, Wen-Bin;Tang, Wei-Xue
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.6
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    • pp.2799-2806
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    • 2012
  • The aim of this study was to elucidate the role of the integrin-linked kinase (ILK) gene in development of human bladder transitional cell carcinoma (BTCC). Expression of ILK protein and ILK mRNA in 56 cases of human BTCC tissue and in 30 cases of adjacent normal bladder tissue was detected by immunohistochemistry S-P and reverse transcription polymerase chain reaction (RT-PCR), respectively. Four specific miRNA RNAi vectors targeting human ILK were synthesized and transfected into BIU-87 cells by liposome to obtain stable expression cell strains. The influence of ILK on proliferation of BTCC was detected by MTT, FCM on athymic mouse tumorigenesis. The positive rate of ILK protein in BTCC tissue (53.6%) was much higher than adjacent normal bladder tissue (10.0%) (p<0.05). Similarly, expression of ILK mRNA in BTCC tissue ($0.540{\pm}0.083$) was significantly higher than in adjacent normal bladder tissue ($0.492{\pm}0.070$) (p<0.05). MTT showed that the proliferation ability of miRNA-ILK transfected group was clearly decreased (p<0.05), the cell cycle being arrested in G0/G1-S, an tumorigenesis in vivo was also significantly reduced (p<0.05). ILK gene transcription and protein expression may be involved in the development of BTCC, so that ILK might be the new marker for early diagnosis and the new target for gene treatment.

The Effects of Jeoreongchajeonja-tang(Zhulingjuqianzi-tang) on the βA and LPS Induced BV2 microglial cell (저령차전자탕(豬苓車前子湯)이 βA와 LPS로 처리된 BV2 microglial cell에 미치는 영향)

  • Ryu, Chang-Hee;Jung, In-Chul;Lee, Sang-Ryong
    • Journal of Oriental Neuropsychiatry
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    • v.23 no.1
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    • pp.145-159
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    • 2012
  • Objectives : This research investigates the effect of the JCT extract regarding Alzheimer's disease. Methods : The effects of the JCT extract on IL-$1{\beta}$, IL-6, TNF-${\alpha}$, COX-2, NOS-II mRNA, APP mRNA, BACE mRNA, Nitric oxide(NO), and ${\beta}A$ protein production in the BV2 microglia cell lines treated with LPS and ${\beta}A$ were investigated. Results : 1. The JCT extract suppressed the expression of IL-$1{\beta}$, IL-6, TNF-${\alpha}$, COX-2, and NOS-II mRNA in BV2 microglial cell line treated with LPS and ${\beta}A$. 2. The JCT extract suppressed the expression of BACE and APP mRNA in BV2 microglial cell line treated with LPS and ${\beta}A$. 3. The JCT extract suppressed the expression of Nitric oxide(NO) in BV2 microglial cell line treated with LPS and ${\beta}A$. 4. The JCT extract suppressed the expression of ${\beta}A$ protein production in BV2 microglial cell line treated with LPS and ${\beta}A$. Conclusions : These results suggest that the JCT group may be effective for the treatment of Alzheimer's disease. Thus, JCT could be considered among the future therapeutic drugs indicated for the treatment of Alzheimer's disease.

16S rRNA Gene Sequence-based Microbial Diversity Analyses of the Geothermal Areas of Cisolok, Kamojang, and Likupang in Indonesia (16S rRNA 분석을 통한 인도네시아의 Cisolok, Kamojang, Likupang 지열지대 내 미생물 다양성 분석)

  • Seo, Myung-Ji;Kim, Jeong-Nyeo;Pyun, Yu-Ryang
    • Microbiology and Biotechnology Letters
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    • v.40 no.3
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    • pp.268-273
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    • 2012
  • Microbial diversity analyses were performed in several geothermal areas in Indonesia using a culture-independent approach with 16S rRNA gene sequencing. All areas and the majority of samples were noted as being affiliated with Proteobacteria. In addition, unclassified bacteria with no phylum affiliation were detected at an incidence rate of 20.0-26.5% in every location. The majority groupings in the geothermal hot stream in Cisolok belonged to ${\beta}$-Proteobacteria (27.1%) and Cyanobacteria (11.0%), whereas the majority from the volcanic area in Kamojang was ${\gamma}$-Proteobacteria (51.5%) followed by Aquificales (12.9%). The predominant groups around an underwater thermal vent in the sea at Likupang were ${\gamma}$-Proteobacteria (33.3%) and then Bacteroidetes (27.6%). This detailed microbial community analyses of each area strongly support a possible association with plausible community groups and environmental habitats, such as extremely geothermal or marine habitats. This study has significantly contributed to the expansion of scientific knowledge of the microbial community in Indonesia.

Molecular Characterization and Survey of the Infection Rate of Orchid fleck virus in Commercial Orchids

  • Kim, Sung-Ryul;Yoon, Ju-Yoon;Choi, Gug-Sun;Chang, Moo-Ung;Choi, Jang-Kyung;Chung, Bong-Nam
    • The Plant Pathology Journal
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    • v.26 no.2
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    • pp.130-138
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    • 2010
  • Orchid fleck virus (OFV) is an unassigned plant virus in the family Rhabdoviridae. OFV was isolated from Cymbidium sp. showing oval necrotic lesions on their leaves in Korea, and designated as OFV-NHHS1. The complete nucleotide sequence of the RNA1 (6,413 nt) (GenBank accession no. AB516442) and RNA2 (6,001 nt) (GenBank accession no. AB516441) was determined in this study. RNA1 and RNA2 contained five and one ORF respectively. RNA1 encodes nucleocapsid (N) of 49 kDa, ORF2 of 26 kDa, ORF3 of 38 kDa, ORF4 of 20 kDa and glycoprotein (G) of 61 kDa proteins, whereas RNA2 encodes a single polymerase of 212 kDa. OFV-NHHS1 shared extremely high similarity of 98.6-100% and 98.9-99.6% in nucleotidle and amino acid sequences with a Japanese isolate, OFV-so, respectively. However, the N, G and L of OFV-NHHS1 revealed 6.9-19.3%, 7.3-12.0%, and 13.4-26.6% identities to those of 29 Rhabdoviruses, respectively. To survey the infection rate of OFV in commercial orchids in Korea, 51 Cymbidium sp., 10 Phalaenopsis sp., 22 Oncidium sp. and 21 Dendrobium sp. plants that showed typical viral symptoms were collected. RT-PCR with specific primers for detection of Cymbidium mosaic virus (CymMV), ORSV and OFV showed high infection rate by ORSV alone and double infection by ORSV and CymMV. One of the orchids tested was infected with OFV. This is the first report of the complete nucleotide sequences of OFV isolated in Korea.

Cosuppression and RNAi induced by Arabidopsis ortholog gene sequences in tobacco

  • Oka, Shin-Ichiro;Midorikawa, Kaoru;Kodama, Hiroaki
    • Plant Biotechnology Reports
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    • v.4 no.3
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    • pp.185-192
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    • 2010
  • The Arabidopsis ${\omega}$-3 fatty acid desaturase (AtFAD7) catalyzes the synthesis of trienoic fatty acids (TA). A transgenic tobacco line, T15, was produced by a sense AtFAD7 construct and showed a cosuppression-like phenotype, namely extremely low TA levels. The sequence similarity between AtFAD7 and a tobacco ortholog gene, NtFAD7, was moderate (about 69%) in the coding sequences. AtFAD7 siRNAs accumulated at a high level, and both AtFAD7 and NtFAD7 mRNAs are degraded in T15 plants. The low-TA phenotype in T15 was dependent on a tobacco RNA-dependent RNA polymerase6 (NtRDR6). We also produced tobacco RNAi plants targeting AtFAD7 gene sequences. The AtFAD7 siRNA level was trace, which was associated with a slight reduction in leaf TA level. Unexpectedly, this RNAi plant showed an increased NtFAD7 transcript level. To investigate the effect of translational inhibition on stability of the NtFAD7 mRNAs, leaves of the wild-type tobacco plants were treated with a translational inhibitor, cycloheximide. The level of NtFAD7 mRNAs significantly increased after cycloheximde treatment. These results suggest that the translational inhibition by low levels of AtFAD7 siRNAs or by cycloheximide increased stability of NtFAD7 mRNA. The degree of silencing by an RNAi construct targeting the AtFAD7 gene was increased by co-existence of the AtFAD7 transgene, where NtRDR6-dependent amplification of siRNAs occurred. These results indicate that NtRDR6 can emphasize silencing effects in both cosuppression and RNAi.