• Title/Summary/Keyword: DNA complex

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Isolation of Nontuberculous Mycobacteria (NTM) from Air Conditioner Dust (에어컨 먼지내 비결핵 항산균의 동정)

  • Choi, Seung Gu;Choi, Myeong Sik
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.4
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    • pp.435-438
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    • 2017
  • Nontuberculous mycobacteria (NTM) contains M. avium-intracellular complex (MAC), M. fortuitum, M. chelonae, M. abscessus, M. kansasii, etc., which causes infections in swine, poultry, and other animals. These bacteria are opportunistic pathogens that can contaminate people, and in doing so, cause various social economic problems. This study proved that NTM are also present in air conditioners of multiuse facilities in frequently visited populated areas. The results of this study are meaningful because they showed that pathogenic microbes can live in the surroundings and cause diseases to people with impaired immunity. This study used the AFB stain, L-J medium culture, and PCR method for the detection of MTB and NTM in air conditioner dust. MTB was not detected in any of the collected samples, while NTM was detected in 2 out of 40 samples (5%). Most people living in the modern environment cannot avoid being exposed to air conditioners so special attention is needed for sanitary inspections and the management of air conditioners. Moreover, it is important to study NTM through various individuals, accumulate the related data and establish the methods and standards of hygiene management system to reduce NTM infections in public areas. Moreover, it would be necessary to identify the precise species of NTM through DNA sequencing and evaluate the effects of NTM on the pulmonary tuberculosis in immunodeficiency patients.

Nucleotide Sequence and Cloning of sfs4, One of the Genes Involved in the CRP-Dependent Expression of E. coli mal Genes. (CRP 의존성 maltose 대사 촉진 유전자 sfs4의 클로닝 및 염기배열 결정)

  • Chung, Soo-Yeol;Cho, Moo-Je;Jeong, Hee-Tae;Choi, Yong-Lark
    • Applied Biological Chemistry
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    • v.38 no.2
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    • pp.111-117
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    • 1995
  • In Escherichia coli, CRP forms a complex with cAMP and acts as a transcriptional regulator of many genes, including sugar metabolism operons. The E. coli MK2001, which is introduced the altered crp, is functional in the expression of lac, ara and man, in the absence of cAMP. However, the expression of mal gene is fully activated by the addition of cAMP or cGMP. The object of the study is cloning of the sfs (sugar fermentation stimulation) genes, which was involved in regulation of mal gene expression with the altered crp gene, and structural analysis and characterization of the genes at the molecular level. We have cloned 5 different E. coli genes which stimulate the maltose metabolism in a crp, cya::km (MK2001) background. Newly identified genes were designated as sfs. One of the sfs genes (pPC1), located at the 53.2 min map position on the E. coli chromosome, was further analyzed. Expression of the genes, which is involved in maltose metabolism, malQ (amylomaltase), was increased to 5.8-fold in the presence of a plasmid, pAP5, containing the subcloned sfs4 gene. The nucleotide seguence of a common 2,126 bp segment of the pPCM1 was determined and two open reading frames (ORF1 and ORF2) were detected. The ORF1 encodes the sfs4 gene and ORF2 encodes a truncated protein. Potential CRP binding site is located in the upstream of the putative promoter in the regulatory region. Expression of the cloned sfs4 gene was positively regulated by the cAMP-CRP complex.

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Novel Molecular-Based Approaches for Analyzing Microbial Diversity in Raw-Milk Long-Ripened Cheeses: A Review (비살균 숙성 치즈의 미생물균총 분석에 이용되는 새롭게 개발된 분자생물학적 방법: 총설)

  • Kim, Dong-Hyeon;Chon, Jung-Whan;Kim, Hyunsook;Lee, Soo-Kyung;Kim, Hong-Seok;Lee, Joo-Yeon;Yim, Jin-Hyuk;Song, Kwang-Young;Kim, Young-Ji;Kang, Il-Byung;Jeong, Dana;Park, Jin-Hyeong;Jang, Ho-Seok;Seo, Kun-Ho
    • Journal of Dairy Science and Biotechnology
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    • v.34 no.1
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    • pp.9-20
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    • 2016
  • Various microflora, including lactic acid bacteria, are important and necessary components of various cheeses and have significant roles in cheese manufacturing and ripening. In general, the starter culture and secondary microflora could affect the physicochemical properties of various cheeses and could contribute to modifications during manufacturing and ripening. Therefore, during cheese manufacturing and ripening, microbial diversity may depend on continuous interactions among microflora and various environmental conditions. The microbial diversity of cheese is very complex and difficult to control using the classical microbiological techniques. However, recent culture-independent methods have been rapidly developed for microflora in cheese, which could be directly detected using DNA (and/or RNA) in combination with culture-dependent methods. Therefore, this review summarizes state-of-the-art molecular methods to analyze microbial communities in order to understand the properties that affect quality and ripening as well as the complex microbial diversity of various raw-milk, long-ripened cheeses.

A new combination for Saxifraga octopetala (Saxifragaceae) and its phylogenetic relationship (구실바위취의 신조합명 및 계통 유연관계)

  • Kim, Yong-In;Cho, Seong Hyun;Kim, Bo-Yun;Lee, Jung-Hoon;Kang, Dae-Hyun;Kim, Soonok;Doudkin, Roman V.;Kim, Young-Dong
    • Korean Journal of Plant Taxonomy
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    • v.45 no.4
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    • pp.306-317
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    • 2015
  • This study was conducted to address the taxonomic status of the Korean endemic species Saxifraga octopetala, which is sometimes considered conspecific to Micranthes manchuriensis. Extensive molecular phylogenetic analysis using nrITS sequences as well as morphological examination of type specimens of the two species were undertaken to ascertain the phylogenetic position and species delimitation of S. octopetala. In the resulting nrITS trees, a total of 65 accessions representing S. octopetala grouped together and nested within the Micranthes clade, exhibiting a close relationship with M. nelsoniana and M. manchuriensis. Multiple accessions of M. manchuriensis collected from China and Russia also formed a clade, showing a sister group relationship with M. nelsoniana var. pacifica and M. fusca. The ambiguous species entity of S. octopetala is thought to have originated from Nakai's misinterpretation of Wilford's collection (type specimens of M. manchuriensis), which is a complex collection including an inflorescence of M. nelsoniana. In spite of apparent morphological similarity between S. octopetala and M. manchuriensis, they differ in the presence and absence of underground stolons. The distinct position of S. octopetala within the Micranthes clade on the nrITS tree suggests that it should retain species status in Micranthes. Thus a new combination (Micranthes octopetala) is proposed.

The Mitochondrial Warburg Effect: A Cancer Enigma

  • Kim, Hans H.;Joo, Hyun;Kim, Tae-Ho;Kim, Eui-Yong;Park, Seok-Ju;Park, Ji-Kyoung;Kim, Han-Jip
    • Interdisciplinary Bio Central
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    • v.1 no.2
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    • pp.7.1-7.7
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    • 2009
  • "To be, or not to be?" This question is not only Hamlet's agony but also the dilemma of mitochondria in a cancer cell. Cancer cells have a high glycolysis rate even in the presence of oxygen. This feature of cancer cells is known as the Warburg effect, named for the first scientist to observe it, Otto Warburg, who assumed that because of mitochondrial malfunction, cancer cells had to depend on anaerobic glycolysis to generate ATP. It was demonstrated, however, that cancer cells with intact mitochondria also showed evidence of the Warburg effect. Thus, an alternative explanation was proposed: the Warburg effect helps cancer cells harness additional ATP to meet the high energy demand required for their extraordinary growth while providing a basic building block of metabolites for their proliferation. A third view suggests that the Warburg effect is a defense mechanism, protecting cancer cells from the higher than usual oxidative environment in which they survive. Interestingly, the latter view does not conflict with the high-energy production view, as increased glucose metabolism enables cancer cells to produce larger amounts of both antioxidants to fight oxidative stress and ATP and metabolites for growth. The combination of these two different hypotheses may explain the Warburg effect, but critical questions at the mechanistic level remain to be explored. Cancer shows complex and multi-faceted behaviors. Previously, there has been no overall plan or systematic approach to integrate and interpret the complex signaling in cancer cells. A new paradigm of collaboration and a well-designed systemic approach will supply answers to fill the gaps in current cancer knowledge and will accelerate the discovery of the connections behind the Warburg mystery. An integrated understanding of cancer complexity and tumorigenesis is necessary to expand the frontiers of cancer cell biology.

Phylogeny of Korean Viola based on ITS sequences (ITS 염기서열에 의한 한국산 제비꽃속(Viola)의 계통 유연관계)

  • Yoo, Ki-Oug;Jang, Su-Kil;Lee, Woo-Tchul
    • Korean Journal of Plant Taxonomy
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    • v.35 no.1
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    • pp.7-23
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    • 2005
  • Molecular phylogenetic studies were conducted to evaluate interspecific relationships in 40 populations of Viola including 35 Korean taxa, four Japanese populations and one outgroup using nuclear ribosomal ITS sequences. The phylogenetic analyses were conducted using parsimony and neighbor-joining methods. Subsection Trigonocarpae of section Nomimium appeared as the most basal clade within the Korean Viola. Section Dischidium and Chamaemelanium was monophyletidbootstrap 100%) and placed between subsect. Trigonocarpae and three other subsections of sect. Nomimium. Sect. Nomimium was paraphyletic. Although each subsectional grouping was in accordance with previous infrageneric classification based on morphological characters, yet discordance remained at the series level. Two evolutionary trends observed in the ITS tree were as follows. First, subsect. Trigonocarpae(x=10) was derived from the outgroup(x=6); Second, subsects. Bilobatae and Vaginatae(x=10 or 12), and subsect. Patellares(x=12) of sect. Nomimium were originated from sects. Dischidium and Chamaemelanium(x=6). Viola albida complex including three very closely related taxa was recognized as independent group within subsect. Patellares in parsimony tree. This result suggested that they should be treated as a taxa in series Pinnatae. Phylogenetic position of a putative hybrid species, Viola woosanensis was not supported with previous morphological hypothesis.

Transcriptomic Analysis of the Difference of Bovine Satellite Cell Between Longissimus dorsi and Semimembranosus on Hanwoo Muscle Tissues (한우의 등심과 사태조직 유래 근육위성세포의 성장단계별 유전발현 차이 분석)

  • Kim, H.J.;Kang, D.H.;Park, B.H.;Lee, W.Y.;Choi, J.H.;Chung, K.Y.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.23 no.1
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    • pp.117-128
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    • 2021
  • The skeletal muscle development of Hanwoo steer has been processed in the prenatal and postnatal periods. Bovine satellite cell located in perimysium of muscle tissues has differentially distributed in peripheral tissues. The study of postnatal development of satellite cells can help understand the genetic and functional regulation of meat characteristics. Factors affecting muscle size increase are related to the accumulation of DNA or synthesis of RNA proteins. In this study, we observed muscle development and differentiation after culturing bovine satellite cells derived from longissimus dorsi and semimembranosus regions of Hanwoo muscle tissue. In addition, RNA sequencing data were analyzed for differentially expressed genes (DEG) involved in intracellular muscle development and growth. The DEG of the two muscle tissues were compared according to 1day, 2day, 4day, and 7day. The overall gene expression level was confirmed by the heat map. Gene Ontology (GO) classification method was used to compare the expression level of gene groups affecting LD and SM development. The histology of GO was consistent with the time-cause change of LD and SM cell morphology. SM showed more active skeletal muscle development than LD. Even within the same time, SM expressed more genes than LD, thus synthesizing more muscle fibers

A Cyclophilin from Griffithsia japonica Has Thermoprotective Activity and Is Affected by CsA

  • Cho, Eun Kyung;Lee, Yoo Kyung;Hong, Choo Bong
    • Molecules and Cells
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    • v.20 no.1
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    • pp.142-150
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    • 2005
  • Members of the multifunctional Cyp family have been isolated from a wide range of organisms. However, few functional studies have been performed on the role of these proteins as chaperones in red alga. For studying the function of cDNA GjCyp-1 isolated from the red alga (Griffithsia japonica), we expressed and purified a recombinant GjCyp-1 containing a hexahistidine tag at the amino-terminus in Escherichia coli. An expressed fusion protein, $H_6GjCyp-1$ maintained the stability of E. coli proteins up to $50^{\circ}C$. For a functional bioassay for recombinant $H_6GjCyp-1$, the viability of E. coli cells overexpressing $H_6GjCyp-1$ was compared with that of cells not expressing $H_6GjCyp-1$ at $50^{\circ}C$. After high temperature treatment for 1 h, E. coli overexpressing $H_6GjCyp-1$ survived about three times longer than E. coli lacking $H_6GjCyp-1$. Measurement of the light scattering of luciferase (luc) showed that GjCyp-1 prevents the aggregation of luc during mild heat stress and that the thermoprotective activity of GjCyp-1 is blocked by cyclosporin A (CsA), an inhibitor of Cyps. Furthermore, the Cyp-CsA complex inhibited the growth of E. coli under normal conditions. The results of the GjCyp-1 bioassays as well as in vitro studies strongly suggest that Cyp confers thermotolerance to E. coli.

Phylogenic Study of Genus Citrus and Two Relative Genera in Korea by trnL-trnF Sequence (trnL-trnF 서열에 의한 한국 귤나무속과 두 근연 식물종의 계통분류학적 연구)

  • Huh, Man-Kyu;Yoon, Hye-Jeong;Choi, Joo-Soo
    • Journal of Life Science
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    • v.21 no.10
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    • pp.1452-1459
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    • 2011
  • Citrus is a common term and genus (Citrus) of flowering plants in the rue family, Rutaceae. Citrus is believed to have originated in the part of Southeast Asia bordered by Northeastern India, Myanmar (Burma) and the Yunnan province of China. The taxonomy and systematics of the genus are complex and the precise number of natural species is unclear, as many of the named species are clonally propagated hybrids, and there is genetic evidence that even some wild, true-breeding species are of a hybrid origin. One of the most popular sequences for phylogenetic inference at the generic and infrageneric levels in plants is the chloroplast trnL-trnF region. We evaluated the seven taxa with the trnL-trnF region to estimate phenotypic relationships within the genera Citrus, Poncirus, and Fortunella in Korea. Alignment of the DNA sequences required the addition of numerous gaps. Sequence variation within Citrus was mostly due to insertion/deletion. Within the genus Citrus, C. lomonia and C. sinensis were relatives and sistered with C. aurantium in the four phylogenetic analyses (MP, ML, ME, and NJ). However, some external nodes were poorly supported.

Diversity of Butyrivibrio Group Bacteria in the Rumen of Goats and Its Response to the Supplementation of Garlic Oil

  • Zhu, Zhi;Hang, Suqin;Mao, Shengyong;Zhu, Weiyun
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.2
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    • pp.179-186
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    • 2014
  • This study aimed to investigate the diversity of the Butyrivibrio group bacteria in goat rumen and its response to garlic oil (GO) supplementation as revealed by molecular analysis of cloned 16S rRNA genes. Six wethers fitted with ruminal fistulas were assigned to two groups for a cross-over design with 28-d experimental period and 14-d interval. Goats were fed a basal diet without (control) or with GO ruminal infusion (0.8 g/d). Ruminal contents were used for DNA extraction collected before morning feeding on d 28. A total bacterial clone library was firstly constructed by nearly full-length 16S rRNA gene cloned sequences using universal primers. The resulting plasmids selected by Butyrivibrio-specific primers were used to construct a Butyrivibrio group-specific bacterial clone library. Butyrivibrio group represented 12.98% and 10.95% of total bacteria in control and GO group, respectively. In libraries, clones were classified to the genus Pseudobutyrivibrio, Butyrivibrio and others within the family Lachnospiraceae. Additionally, some specific clones were observed in GO group, being classified to the genus Ruminococcus and others within the family Ruminococcaceae. Based on the criterion that the similarity was 97% or greater with database sequences, there were 29.73% and 18.42% of clones identified as known isolates (i.e. B. proteoclasticus and Ps. ruminis) in control and GO groups, respectively. Further clones identified as B. fibrisolvens (5.41%) and R. flavefaciens (7.89%) were specifically found in control and GO groups, respectively. The majority of clones resembled Ps. ruminis (98% to 99% similarity), except for Lachnospiraceae bacteria (87% to 92% similarity) in the two libraries. The two clone libraries also appeared different in Shannon diversity index (control 2.47 and GO group 2.91). Our results indicated that the Butyrivibrio group bacteria had a complex community with considerable unknown species in the goat rumen.