• Title/Summary/Keyword: genetic characterization

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Purification and Characterization of a Thermostable ${\beta}-Glycosidase$ from Thermus caldophilus GK24

  • Yoo, Jin-Sang;Han, Ki-Woong;Kim, Hyun-Kyu;Kim, Min-Hong;Kwon, Suk-Tae
    • Journal of Microbiology and Biotechnology
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    • v.10 no.5
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    • pp.638-642
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    • 2000
  • A ${\beta}-glycosidase$ enzyme with $\beta$-D-fucosidase, ${\beta}-D-galactosidase$, and $\beta$-D-glucosidase activities has been purified from Thermus caldophilus GK24. The enzyme was monomeric with a molecular mass of 49 kDa, as evidenced by SDS-PAGE. The $K_m$ values for p-nitrophenyl ${\beta}-D-fucopyranoside$ (p-NPFuc), p-nitrophenyl ${\beta}-D-galactopyranoside$ (p-NPGal), and p-nitrophenyl ${\beta}-D-glucopyranoside$ (p-NPGlu) were 0.23 mM, 6.25 mM, and 0.28 mM, respectively. The enzyme showed optimal pH ranging between 5.5-6.5 and maximum temperature in the range of $85-90^{\circ}C$ for all the above mentioned activities. The half-life of the enzyme in sodium phosphate buffer (pH 6.0) at $80^{\circ}C$ was approximately 7 h. The p-NPGal hydrolyzing activity of Tca ${\beta}-glycosidase$ was strongly activated by L-histidine, while the p-NPFuc and p-NPGlu hydrolyzing activities of Tca ${\beta}-glycosidase$ were not affected at all by the amino acid. These results suggest differences in the conformation or in the reactive residues at the active site of Tca ${\beta}-glycosidase$.

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Genetic Characterization of Encephalomyocarditis Virus Isolated from Aborted Swine Fetus in Korea

  • Song, Min-Suk;Joo, Young-Ho;Lee, Eun-Ho;Shin, Jin-Young;Kim, Chul-Jung;Shin, Kwang-Soon;Sung, Moon-Hee;Choi, Young-Ki
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1570-1576
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    • 2006
  • An encephalomyocarditis virus (EMCV-CBNU) was isolated from an aborted swine fetus in October 2005. To investigate the genetic origin and virulence of the EMCV-CBNU strain, we determined the complete sequence of the virus and tested its virulence in mice. Genetic characterization revealed that the RNA genome was composed of 7,713 nucleotides with a single open reading frame (2,292 amino acids), coding 12 proteins. The EMCV-CBNU had the shortest poly(C) tract, consisting of 10 C's ($C_{10}$), compared with all the other EMCV strains reported in GenBank. Amino acid and phylogenetic analyses showed that EMCV-CBNU had the highest genetic identity with strain 2887A (99.7%), which was originally isolated from a fetus in a pig breeding farm that had a history of reproductive failure. Because rodents are the natural host of EMCV, we investigated the virulence of EMCV-CBNU in mice. Surprisingly, all mice inoculated with more than $1{\times}10^2\;TCID_{50}/0.1ml$ of EMCV-CBNU showed symptoms of hind limb paralysis and eventually died during 3 and 8 days postinoculation (DPI). Furthermore, when we inoculated the virus into pregnant mice, all dams and their fetuses died in 6 DPI. This is the first report on a full genomic analysis of swine EMCV in Korea, which exhibits high virulence in mice.

Analysis of Genetic Diversity and Relationships of Korean Native Black Goat using Microsatellite Markers (초위성체 마커를 이용한 한국 재래 흑염소의 유전적 다양성 및 유연관계 분석)

  • Park, Byeong Kyu;Kim, Yi Seul;Seong, Jiyeon;Kong, Hong Sik
    • Journal of Animal Reproduction and Biotechnology
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    • v.34 no.3
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    • pp.183-189
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    • 2019
  • The aim of this study was to assess the levels of genetic diversity and relationships of Korean native black goat (n = 58), compared with the exotic breed, Boer (n = 97). For the analysis of genetic characterization 11 microsatellite markers (MAF065, INRA063, CSRD247, OarFCB20, SRCRSP5, INRA006, ILSTS008, ILSTS011, INRA005, ILSTS087, SRCRSP8) were genotyped. The number of alleles was observed 3 (INRA005) to 10 (SRCRSP8) each markers. The mean expected and observed heterozygosity (Hexp and Hobs) and polymorphism information content (PIC) for the Korean native black goat breed varied from 0.551 to 0.860, 0.517 to 0.948 and 0.464 to 0.835, respectively. Principal Components Analysis (PCoA) and FCA results showed that Korean native black goat breed was confirmed to be clearly separated from bore breed. These results were scientific evidence that Korean native black goat represents a unique and valuable animal genetic resource.

Peroxiredoxin(PRX) gene family characterization in aves

  • Shin, Sang-Su;Kim, Tae-Min;Shin, Ji-Hye;Park, Tae-Sub;Kim, Jin-Kyoo;Kim, Hee-Bal;Han, Jae-Yong
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2004.11a
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    • pp.11-12
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    • 2004
  • Peroxiredoxin(PRX)은 원핵세포에서 진핵세포에 이르기까지 세포 내부적으로 발생된 과산화물로부터 자신을 보호하는 중요한 항산화단백질이다. 포유류에서는 아직까지 여섯 개의 다른 동형체가 밝혀졌으며, 조류에서는 아직 발표된 바가 없다. 이 실험을 통해 최초로 조류의 PRX 단백질군들의 특성을 분석하였다. 생물정보분석기법을 통해 알아본 결과, 조류에서는 최소한 진화적으로 보존된 4개의 다른 PRX 단백질로 구성됨을 알 수 있다. 또한 닭의 PRXs로 in vitro 실험을 진행한 결과, 포유류의 것과 비슷한 항산화 활성을 나타냄을 알 수 있었다. 닭의 PRX는 조직 비특이적으로 발현하였으며, 이는 항산화 물질의 피해로부터 모든 조직을 보호하기 위한 필수적 요소이기 때문일 것으로 추정된다. 결론적으로, 생물정보분석기법을 통하여 추정할 수 있는 닭의 기능성 유전자군을 효과적으로 찾을 수 있고, in vitro 실험을 통하여 그 기능을 확인할 수 있었다.

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Production and Characterization of Monoclonal Antibodies to Bovine Brain Succinic Semialdehyde Reductase

  • Park, E.Y.;Park, S.Y.;Jang, S.H.;Song, M.S.;Cho, S.W.;Park, S.Y.
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1995.04a
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    • pp.72-72
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    • 1995
  • Monoclonal antibodies against bovine brain succinic semialdehyde reductase were produced and characterized. A total of nine monoclonal antibodies recognizing different epitopes of the enzyme were obtained, of which two inhibited the enzyme activity and three stained cytosol of rat spinal cord neurons as observed by indirect immunofluorescence microscopy. When unfractionated total proteins of bovine brain homogenate were seperated by gel electrophoresis and immunoblotted, the antibodies specifically recognized a single protein band of 34 kDa, which comigrates with purified bovine succinic semialdehyde reducatase Using the antisuccinic semialdehyde reductase antibodies as probes, we investigated the cross-reactivites of brain succinic semialdehyde reductases from some mammalian and an avian species. The immunoreactive bands on Western blots appeared to be the same in molecular mass-34 kDa-in all animal species tested, including humans. The result indicated that brain succinic semialdehyde reductase is distinct from other aldehyde reductases and that mammalian brains contain only one succinic semialdehyde reductase. Moreover, the enzymes among the species are imunologically very similar, although some properties of the enzymes reported previously were different from one another.

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Studies on Genetic Diversity of Buckwheat Germplasms

  • Gao, Xiu-Dong;Kim, Jae-Hak;Park, Cheol-Ho;Hong, Soon-Kwan
    • Korean Journal of Plant Resources
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    • v.23 no.3
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    • pp.214-222
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    • 2010
  • Many research results have indicated that many kinds of useful ingredients are rich in buckwheat, which have high nutritional values and medicinal properties, so, buckwheat has been cultivated around Asia and Europe. In this paper, genetic diversity of common and tartary buckwheat germplasms were studied based on morphological and molecular markers in order to provide useful information for conservation and utilization of buckwheat genetic resources. The length, width, thick, value and weight of the seed of the common and tartary buckwheat were measured and analyzed by the statistics methods. The result has shown that there are morphological variation both in common buckwheat and tartary buckwheat seeds collected from different regions. It also has shown that the morphological variation of tartary seeds was significantly correlated to geographical regions. The nuclear ribosomal internal transcribed spacers (ITS) of the tartary buckwheat collected from different countries were cloned, sequenced and statistically analyzed. The data showed that ITS sequences were informative to analyze the phylogeny of the tartary buckwheat and the data also showed that the genetic distances varied among different tartary buckwheat seeds collected from different countries.

Isolation and Characterization of Malate Dehydrogenase Gene from Panax ginseng C.A. Meyer (고려인삼에서 Malate Dehydrogenase 유전자의 분리 및 분석)

  • Kim, Yu-Jin;Shim, Ju-Sun;Lee, Jung-Hye;Jung, Dae-Young;In, Jun-Gyo;Lee, Bum-Soo;Min, Byung-Hoon;Yang, Deok-Chun
    • Korean Journal of Medicinal Crop Science
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    • v.16 no.4
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    • pp.261-267
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    • 2008
  • Malate dehydrogenase is a ubiquitous enzyme in plants, involving in a range of metabolic processes depending on its subcellular location. A malate dehydrogenase (PgMDH) cDNA was isolated and characterized from the root of Panax ginseng C. A. Meyer. The deduced amino acid sequence of PgMDH showed high similarity with the NAD-dependent mitochondrial malate dehydrogenase from Glycinemax (P17783), Eucalyptus gunnii (P46487), and Lycopersicon esculentum (AAU29198). And the segment of a malate dehydrogenase gene was amplified through RT-PCR. The expression of PgMDH was increased after treatments of chilling, salt, UV, cadmium or copper treatment.

Characterization and biovar. cetermination of agrobacterium tumefaciens T7 isolated in Korea (한국에서 분리한 agrobacterium tumefaciens T7의 특성과 biovar.결정)

  • Rhee, Y.;Kim, C. J.;Kim, S. H.;Yoo, I. D.;Mheen, T. I.
    • Korean Journal of Microbiology
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    • v.25 no.1
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    • pp.17-22
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    • 1987
  • For the purpose of securing of strains which can be usefully utilized to study symbiosis between Rhizobium and legume plant, A. tumefaciens T7 was isolated and characterized and then subgroup biovar was determined. A. tumefaciens T7 induced smooth tumor like nopaline type one and did not grow at $37^{\circ}C$ and in the presence of 2% NaCl on yeast extract mannitol medium. The strain was able to grow on the New and Kerr selective media and utilize erythritol but not phenylalanine, tryptophan, and tartarate as a sole carbon source. Negative results were obtained from 3-keto-lactose production and oxidase test. The strain produced alkalifrom malonate and citrate and showed acid litmus milk reaction At least two large plasmids were detected in the cell lysate. According to all of these results, it could be concluded that subdivision of isolated strain was biovar 2.

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Morphological and Genetic Characterization of Caffeine-Rich and -Poor Tea Tree (Camellia sinensis L.) Lines

  • Kim, Yong-Duck;Jeong, Mi-Jin;Song, Hyun-Jin;Yun, Seok-Rak;Heo, Chang-Mi;Kim, Chang-Soo;Moon, Hyun-Shik;Choi, Myung-Suk
    • Journal of agriculture & life science
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    • v.45 no.5
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    • pp.1-8
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    • 2011
  • In this study, 160 tea tree (Camellia sinensis L.) lines were classified by caffeine content using colorimetric methods. Among them, caffeine-rich lines (HR-78, HR-137, HR-82 and HR-123) and poor lines (HP-85, HP-88, HP-19, and HP-131) were selected. To know the difference in morphological and genetic characters between caffeine-rich and poor lines, we used leaf/shoot growth and RAPD methods. Cluster pattern of morphological characters (leaf width, leaf length, leaf area and shoot length) showed that shoot length was longer in caffein-rich lines than in -poor lines. In genetic analysis, amplified DNA bands having various sizes were detected in RAPD analysis where 30 random primers were used. However, the discriminated primer set that distinguish caffein-rich tree line from -poor lines was not found. These results can be used as the basic data to determine the morphological and genetic differences among caffein-rich and -poor lines.

Morphological and molecular analysis of indigenous Myanmar mango (Mangifera indica L.) landraces around Kyaukse district

  • Kyaing, May Sandar;Soe, April Nwet Yee;Myint, Moe Moe;Htway, Honey Thet Paing;Yi, Khin Pyone;Phyo, Seinn Sandar May;Hlaing, Nwe Nwe Soe
    • Journal of Plant Biotechnology
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    • v.46 no.2
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    • pp.61-70
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    • 2019
  • There is vast genetic diversity of Myanmar Mangoes. This study mainly focused on indigenous thirteen different mango landraces cultivated in central area of Myanmar, Kyauk-se District and their fruit characteristics by 18 descriptors together with genetic relationship among them by 12 SSR markers. Based on the morpho-physical characters, a wide variation among accessions was found. Genetic characterization of thirteen mango genotypes resulted in the detection of 302 scorable polymorphic bands with an average of 4.33 alleles per locus and an average polymorphism information content (PIC) of 0.7. All the genotypes were grouped into two major clusters by UPGMA cluster analysis and a genetic similarity was observed in a range of 61 ~ 85%. This study may somehow contribute insights into the identification of regional mango diversity in Myanmar and would be useful for future mango breeding program.