• Title/Summary/Keyword: Positive sequence

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Cloning, Sequencing and Expression of the Gene Encoding a Thermostable β-Xylosidase from Paenibacillus sp. DG-22 (Paenibacillus sp. DG-22로부터 열에 안정한 β-xylosidase를 암호화하는 유전자의 클로닝, 염기서열결정 및 발현)

  • Lee, Tae-Hyeong;Lee, Yong-Eok
    • Journal of Life Science
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    • v.17 no.9 s.89
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    • pp.1197-1203
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    • 2007
  • A genomic DNA library of the bacterium Paenibacillus sp. DG-22 was constructed and the ${\beta}-xylosi-dase-positive$ clones were identified using the fluorogenic substrate $4-methylumbelliferyl-{\beta}-D-xylopyr-anoside$ $({\beta}MUX)$. A recombinant plasmid was isolated from the clone and 4.3-kb inserted DNA was sequenced. The ${\beta}-xylosidase$ gene (xylA) was comprised of a 2,106 bp open reading frame (ORF) en-coding 701 amino acids with a molecular weight of 78,710 dalton and a pI of 5.0. The deduced amino acid sequence of the xylA gene product had significant similarity with ${\beta}-xylosidases$ classified into family 52 of glycosyl hydrolases. The xylA gene was subcloned into the pQE60 expression vector to fuse with six histidine-tag. The recombinant ${\beta}-xylosidase$ $(XylA-H_6)$ was purified to homogeneity by heat-treatment and immobilized metal affinity chromatography. The pH and temperature optima of the $XylA-H_6$ enzyme were pH 5.5-6.0 and $60^{\circ}C$, respectively.

Strategic Approach of Dredging the Port of Busan with the Change of Global Port Logistics Circumstances: Focusing on Financial Validity (항만물류환경 변화에 따른 부산항의 증심 준설 전략 - 재무적 타당성을 중심으로 -)

  • Ha, Myung-Shin;Chang, Byoung-Ky
    • Journal of Korea Port Economic Association
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    • v.24 no.2
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    • pp.131-154
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    • 2008
  • The purpose of this paper is to verify the financial validity of dredging the port of Busan with the change of global port logistics circumstances. In order to do so, NPV, IRR, B/C ratio, DPP techniques are adopted. The whole dredging at Northern port area is supposed to be negative at the depth of 16m water. Based upon its financial validity, however, the priority is Shinsundae (option 1) followed by Shinsundae (option 1) and Gamman (option 1), Shinsundae (option 1) and Gamman (option 2), Shinsundae (option 2) and Gamman (option 2), Shinsundae (option 2) and Gamman (option 2) and ShinGamman in sequence. In addition to this, Busan New port is considered to be the positive financial validity at the dredging of 16m water depth. This is because the ultra containerships of over 8,000TEU are trying to call at the port and Hanjin shipping and Hyundai Merchant shipping companies which are now the most significant customers in Northern port are scheduled to divert it.

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Inheritance and Heritability of Telomere Length in Chicken (닭 텔로미어 길이의 유전력 추정과 유전 전이 양상)

  • Park, Dan Bi;Sohn, Sea Hwan
    • Korean Journal of Poultry Science
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    • v.41 no.3
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    • pp.217-225
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    • 2014
  • Telomeres are the ends of the eukaryotic chromosomes and consist of a tandem repetitive DNA sequence and shelterin protein complex. The function of telomere is to protect chromosome. Telomere length in somatic cells tends to decrease with organismal age due to the end replication problem. However, several factors at the genetic, epigenetic and environmental level affect telomere length. In this study, we estimated heritability of telomere length and investigated inheritance of telomeres in a chicken. Telomere length of lymphocytes was analyzed by semi-quantitative polymerase chain reaction using telomere primer and quantitative fluorescence in situ hybridization using telomeric DNA probe. In results, heritability of telomere length was estimated 0.9 at birth by offspring-parent regression analysis and was estimated 0.03 and 0.04 at 10 and 30 weeks old, respectively, by parental variance analysis. There was a significant positive correlation in telomere length between father and their offspring (r=0.348), and mother and their offspring (r=0.380). In inheritance patterns of telomere length, the influence of paternal and maternal effect on their offspring was similar. The influence of inherited telomeres on male and female progeny was also roughly alike. These results implicated that imprinting of parental telomere length was regulated by autosomal genes, not sex linked genes. In addition, telomere length of offspring at birth did not differ along with their maternal age. Thus, maternal age does not affects telomere length in their offspring at birth owing to cellular reprogramming at early embryonic stage.

Diversity of vir Genes in Plasmodium vivax from Endemic Regions in the Republic of Korea: an Initial Evaluation

  • Son, Ui-han;Dinzouna-Boutamba, Sylvatrie-Danne;Lee, Sanghyun;Yun, Hae Soo;Kim, Jung-Yeon;Joo, So-Young;Jeong, Sookwan;Rhee, Man Hee;Hong, Yeonchul;Chung, Dong-Il;Kwak, Dongmi;Goo, Youn-Kyoung
    • Parasites, Hosts and Diseases
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    • v.55 no.2
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    • pp.149-158
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    • 2017
  • Variant surface antigens (VSAs) encoded by pir families are considered to be the key proteins used by many Plasmodium spp. to escape the host immune system by antigenic variation. This attribute of VSAs is a critical issue in the development of a novel vaccine. In this regard, a population genetic study of vir genes from Plasmodium vivax was performed in the Republic of Korea (ROK). Eighty-five venous blood samples and 4 of the vir genes, namely vir 27, vir 21, vir 12, and vir 4, were selected for study. The number of segregating sites (S), number of haplotypes (H), haplotype diversity (Hd), DNA diversity (${\pi}$ and ${\Theta}_w$), and Tajima's D test value were conducted. Phylogenetic trees of each gene were constructed. The vir 21 (S=143, H=22, Hd=0.827) was the most genetically diverse gene, and the vir 4 (S=6, H=4, Hd=0.556) was the opposite one. Tajima's D values for vir 27 (1.08530, P>0.1), vir 12 (2.89007, P<0.01), and vir 21 (0.40782, P>0.1) were positive, and that of vir 4 (-1.32162, P>0.1) was negative. All phylogenetic trees showed 2 clades with no particular branching according to the geographical differences and cluster. This study is the first survey on the vir genes in ROK, providing information on the genetic level. The sample sequences from vir 4 showed a clear difference to the Sal-1 reference gene sequence, whereas they were very similar to those from Indian isolates.

Distribution of Human Rotavirus Genotypes in a Tertiary Hospital, Seoul, Korea During 2009-2013 (2009년부터 2013년까지 서울의 일개 대학병원에서 동정된 로타바이러스 유전형의 분포)

  • Han, Tae Hee;Park, Sang-Hun;Chung, Ju-Young;Hwang, Eung-Soo
    • Pediatric Infection and Vaccine
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    • v.22 no.2
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    • pp.81-90
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    • 2015
  • Purpose: Group A rotavirus (RV) is most common etiologic agent of acute gastroenteritis (AGE) in children worldwide. Recently, vaccination has been introduced in several countries to reduce the disease burden caused by RV infections, but continuous surveillance of RV strains is necessary to detect the emergence of potential variants induced by vaccine-immune pressure. This study aimed to investigate the changing pattern of RV genotypes in children with AGE, following the introduction of vaccination in Korea. Methods: Genotyping of RVs by RT-PCR on the basis of VP7 and VP4 gene segment sequence was carried out on 201 rotavirus-positive stool samples, from children hospitalized with AGE between August 2009 and June 2013. We have directly sequenced PCR products and analyzed the phylogenetic tree. Results: The most prevalent G genotype was G9 (33.3%), followed by G1 (22.4%), G3 (15.9%), G2 (6.0%), G4 (3.0%), G10 (1.5%), and mixed G-type (15.4%), with some nontypeable cases (2.5%). The detected P genotypes were P[4] (45.3%), P[8] (43.8%), mixed P-type (10.4 %), and P[2] (0.5%). The G9P[4] genotype was predominantly observed in hospitalized cases in Seoul in 2010/2011, however G1P[8] has been re-emerged as the predominant genotype in the following season (P =0.004). Conclusions: It seems that the periodic fluctuation in predominance of the G1, G3, and G9 strains occurred in Korea during 2009-2013, following the introduction of RV vaccination.

Antifungal Activity of Lactobacillus plantarum Isolated from Kimchi (김치로부터 항진균 활성 Lactobacillus plantarum의 분리 및 특성 규명)

  • Yang, Eun-Ju;Chang, Hae-Choon
    • Microbiology and Biotechnology Letters
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    • v.36 no.4
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    • pp.276-284
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    • 2008
  • A lactic acid bacterium having antifungal activity was isolated from kimchi. It was identified as Lactobacillus plantarum based on its morphological and biochemical properties, and 16S rRNA sequence, and designated as Lb. plantarum AF1. This isolate inhibited the growth of Aspergillus flavus ATCC 22546, A. fumigatus ATCC 96918, A. petrakii PF-1, A. ochraceus PF-2, A. nidulans PF-3, Epicoccum nigrum KF-1, and Cladosporium gossypiicola KF-2 under a dual culture overlay assay. Also, the antimicrobial activity was found to be active against various species of Gram-positive and Gram-negative bacteria. The antifungal activity was found to be stable after heat ($121^{\circ}C$, 15 min) and proteolytic enzyme treatment, but it was unstable over pH 5.0. The antifungal compound(s) was estimated to have a low molecular mass (below 3,000 Da).

Selection of the Auxin and ACC Deaminase Producing Plant Growth Promoting Rhizobacteria from the Coastal Sand Dune Plants (Auxin과 ACC Deaminase를 생산하는 사구식물 복원용 근권세균의 선발)

  • Lim, Jong-Hui;Kim, Jong-Guk;Kim, Sang-Dal
    • Microbiology and Biotechnology Letters
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    • v.36 no.4
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    • pp.268-275
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    • 2008
  • In order to develop the multi-functional rhizobacteria that can exert positive effect on the growth of plants growing in the coastal sand dune located along East Coast of Korea, rhizospheral bacteria of 11 different plants from this area were isolated 1,330 rhizobacteria. Among these, 23 strains were able to produce auxin and had spectrum of antagonism toward various phytopathogenic microbes. To know the mechanism of this antifungal activity, these 23 strains were subjected to further analyses; 19 strains of these produced siderophore as determined by color reaction on CAS-blue plate, 4 strains produced antifungal cellulase as judged by color change on CMC-Congo red plate, 17 strains were able to utilized insoluble phosphate salts, also determined by clear zone formation on PVK medium. Identification of the strain was assigned to all 23 strains by l6s rDNA sequence analysed, and all were identified to be in the genus of Bacillus and Pseudomonas. One strain of these, denoted Pseudomonas fluorescens IB4-14, showed ACC deaminase activity which is known to be involved in the resistance of environmental stress such as salt and drought. Also, P. fluorescens IB4-l4 showed the germination stimulation and roots growth promoting activity on the in vivo assay of Lysimachia mauritiana Lam. (spoonleaf yellow loosestrife).

Isolation and Characterization of Actinomycete Strain BK185 Possessing Antifungal Activity against Ginseng Root Rot Pathogens (인삼 뿌리썩음병균에 항균활성이 있는 방선균 BK185의 분리 및 특성)

  • Kim, Byung-Yong;Bae, Mun-Hyung;Ahn, Jae-Hyung;Weon, Hang-Yeon;Kim, Sung-Il;Kim, Wan-Kyu;Oh, Dong-Chan;Song, Jaekyeong
    • The Korean Journal of Pesticide Science
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    • v.18 no.4
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    • pp.396-403
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    • 2014
  • Ginseng (Panax ginseng C. A. Meyer) is an economically valuable pharmaceutical crop in Korea. In order to find promising biocontrol agents for soil-borne fungal pathogens which infect ginseng roots, we have isolated actinomycete, BK185 from soil. The isolate was investigated for the antifungal activity against to ginseng rot pathogens prior to testing genetic and chemical properties. The strain was identified as Streptomyces sp. using phylogenetic analysis based on 16S rRNA gene sequence. The most closely related species was S. sporoclivatus and S. geldanamycininus with high similarities (>99%). The isolate, BK185 showed positive reaction for PCR detection targeting biosynthetic gene clusters of PKS (Type-I polyketide synthase) and NRPS (Non-ribosomal polypeptide synthetase) genes. Major metabolite from the BK185 was analyzed by The LC/MS and identified to geldamycin, which was known to contained broad antibacterial, antifungal or anticancer activities. The results provide evidences that the strain, BK185 can be promising biocontrol agent for ginseng organic farming.

Isolation and identification of a tricin 4"-O-(threo-β-guaiacylglyceryl) ether producing microorganism from germinated rice (발아 벼로부터 tricin 4"-O-(threo-β-guaiacylglyceryl) ether 생성균주의 분리 및 동정)

  • Yoon, Nara;Jang, Gwi Yeong;Lee, Yoon Jeong;Li, Meishan;Kim, Min Young;Kim, Hyun Young;Lee, Junsoo;Jeong, Heon Sang
    • Korean Journal of Food Science and Technology
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    • v.48 no.4
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    • pp.361-365
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    • 2016
  • This study was conducted to isolate and identify a microorganism that increases tricin-O-(threo-${\beta}$-guaiacylglyceryl) ether (TTGE) content in the hulls of rice (Oryza sativa L.). Bacteria from germinated rice were isolated by enrichment cultivation using yeast mold, luria bertani, potato dextrose and mannitol egg york polymyxin broths. The highest increase in TTGE content ($339.30{\mu}g/g$) was achieved by a microorganism isolated by PDA enrichment cultivation. On the basis of 16S RNA sequence homology and phylogenetic analysis, the isolated bacterium was identified to have 100% similarity with Burkholderia vietnamiensis. The isolated bacteria were short rods, negative for the Gram stain, and positive for the catalase test. The highest TTGE level was $435.86{\mu}g/g$ in 72-h fermented samples, representing a 2.5x increase compared with the control ($175.65{\mu}g/g$). In conclusion, the bacterium isolated from germinated rice extract was Burkholderia vietnamiensis, and the optimum fermentation period to maximize TTGE levels was 72 h. These findings might help in developing functional materials using rice hulls, a waste product of rice milling.

The Utility of TAR Vectors Used for Selective Gene Isolation by TAR Cloning. (TAR Cloning에 의한 선별적 유전자 분리에 사용되는 TAR Vectors의 유용성에 관한 연구)

  • 박정은;이윤주;정윤희;김재우;김승일;김수현;박인호;선우양일;임선희
    • Microbiology and Biotechnology Letters
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    • v.31 no.4
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    • pp.322-328
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    • 2003
  • The Transformation-Associated Recombination (TAR) cloning technique allows selective isolation of chromosomal regions and genes from complex genomes. The procedure requires knowledge of relatively small genomic sequences that reside adjacent to the chromosomal region of interest. This technique involves homologous recombination during yeast spheroplast transformation between genomic DNA and a TAR vector that has 5'and 3' gene targeting sequences. In this study, we examined the minimum size of specific hooks required for a single-copy gene isolation and compared the utility of different TAR vectors, radial and unique vectors, by cloning the same single-copy gene. The efficiency of TAR cloning of the hHPRT gene was same using hooks varying from 750 to 63 bp. The number of transformants decreased approximately 20-fold when the TAR vector contained two unique hooks versus using a radial vector, but the percentage of positive recombinants increased over 2-fold when a unique TAR vector was used. Therefore, we suggest that the two-unique TAR vector is suitable for general TAR cloning given its high selectivity, and the radial TAR vector is more suitable when genomic DNA is in limited quantity, for example, DNA isolated from pathological specimens. Moreover, we confirm the minimal length of a unique sequence in a TAR vector is approximately 60 bp for a single-copy gene isolation.