• Title/Summary/Keyword: Molecular genetic characteristics

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Sequence analysis of ORF4 gene of porcine reproductive and respiratory syndrome virus (PRRSV) Korean isolate CNV-1

  • Park, Jee-yong;Lim, Bae-keun;Kim, Hyun-soo
    • Korean Journal of Veterinary Research
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    • v.39 no.2
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    • pp.294-300
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    • 1999
  • In this study PRRSV was isolated from serum of an infected pig and designated as CNV-1, ORF4 gene was sequenced, and the nucleotide sequence, deduced amino acid sequence and the amino acid sequence of the neutralizing domain was compared with other PRRSV Strains. ORF4 gene of the Korean isolate PRRSV CNV-1 was shown to be 537bp in length, which is the same as US strain ISU55 but 21bp longer than another US strain MN1b, and 15bp shorter than European strain LV. The homologies of the nucleotide sequences between the Korean isolate CNV-1 and the US strains ISU55, MN1b and European strain LV were 91.8%, 88.1%, 67.6%, respectively, and the homologies of the deduced amino acid sequences were 94.4%, 84.4%, 68.5%, respectively. The neutralizing domain of the CNV-1 was shown to be 36 amino acids in length which is the same as ISU55, MN1b, but 4 amino acids shorter than that of the neutralizing domain reported in LV. The homologies of the amino acid sequences of the neutralizing domain between the Korean isolate CNV-1 and the US strains ISU55, MN1b and European strain LV were 92.5%, 85%, 57.5%, respectively. The molecular characteristics of ORF4 gene of the Korean isolate PRRSV CNV-1 shown in this study suggests that the CNV-1 is genetically closer to the US strains. Also the wide variation of the neutralizing domain between the CNV-1 and LV suggests that there is substantial immunogenic variation between the two strains.

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Antiepileptical Properties Of Ginsenosides From Korean Red Ginseng And Ginseng Cell Culture (Dan25)

  • ChepurnovS.A.;Park, Jin-Kyu;vanLuijtelaarE.L.J.M;ChepurnovaN.E.;StrogovS.E.;MikhaylovaO.M.;ArtukhovaM.V.;BerdievR.K.;GoncharovO.B.;SergeevV.I.;TolamachevaE.A.
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2000.10a
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    • pp.116-122
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    • 2000
  • The molecular modification of antiepileptic drugs and direct synthesis of new drugs with the predetermined antiepileptic properties are perspective. New neurochemical attacking to solve the problem including prevention and inhibition of seizures seems to be related to ginsenosides and ginseng polypeptides. The main study based on the severity of febrile convulsions of rat pups has been done from the earlier investigations of antiepileptical action of ginsenosides between KGTRI and MSU (Chepurnov, Park et al., 1995) with different kinds of experimental models of epilepsy. From the cultured cell line DAN25 of ginseng root, the extracts of ginsenosides made in "BIOKHIMMASH" were studied by the project of preclinical anticonvulsant screening (Stables, Kupferberg, 1997). The inhibition of severity of convulsions, decrease of seizures threshold, decrease of audiogenic seizures in rats of different strains and normalization of cerebral blood flow (measured by hydrogen test) were demonstrated in rats after i.c.v., intraperitoneally and orally administration, respectively. The antiepileptical effects by the combination of compounds from ginseng; were compared with the iuluence of Rg1, Rb1, Rc and with the well known antiepileptical drugs such as carbamazepine, valproic acid. The base for the research is obtained by using the WAG/Rij strain (Luijtelaar, Coenen, Kuznetcova), an excellent genetic model for human generalized absence epilepsy. The improving action of gensinosides was effectively demonstrated on the model of electrical kindling of amygdala of WAG/Rij rats with genetically determined absences, and the influences of ginsenosides on the slow wave discharges have also been being investigated. The different characteristics of a kindling process exerted in the sex-different region of the amygdala and demonstrated that the level of sex steroids and content of neurosteroids in amygdaloid tissue can modify the development of seizures. The chemical structures of ginsenosides not only have some principal differences from well-known antiepileptical drugs but the Plant Pharmacology gives us unique possibility to develop new class of antiepileptic drugs and to improve its biological activity.

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One unusual species, Coilia sp. (Engraulidae, Pisces) from the Yellow Sea

  • Kwun, Hyuck-Joon;Kim, Yeong-Hye;Kim, Jong-Bin;Jeong, Choong-Hoon;Kim, Jin-Koo
    • Animal cells and systems
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    • v.14 no.2
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    • pp.137-145
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    • 2010
  • Four specimens of unknown Coilia sp. were collected for the first time from the Yellow Sea in 2008 and compared with Coilia mystus and Coilia nasus. Coilia sp. showed similar morphology to C. mystus and C. nasus, but differed in that its tail was considerably shorter. We conducted an analysis of the morphological and genetic characteristics in an effort to clarify the taxonomic position of Coilia sp. In counts and measurements, Coilia sp. were well distinguished from C. nasus by the number of scutes (42-44 in Coilia sp. vs. 40-45 in C. mystus vs. 45-55 in C. nasus), ratio of dorsal base length to head length (43.4-47.6 vs. 37.9-47.6 vs. 33.0-41.0), and eye length to head length (19.2-20.8 vs. 17.0-22.4 vs. 13.8-18.2). In caudal skeleton of Coilia sp., urostyle, hypural and epural bones were not observed; instead of them, caudal fin rays were supported by the last vertebra, neural and haemal spines' extension. The molecular phylogenetic relationship was analyzed using 414 base-pair 12S rRNA mitochondrial DNA sequences. The Kimura-2-parameter distance between Coilia sp. and C. mystus was 0.3%, but was 1.3% between Coilia sp. and C. nasus. Both the neighbor-joining tree and maximum-likelihood tree showed that Coilia sp. are closely clustered with C. mystus. Therefore, our results suggest that the Coilia sp. may be a deformed fish of C. mystus.

Genetic Evolution and Characteristics of Ichthyotoxic Cochlodinium polykrikoides(Gymnodiniales, Dinophyceae) (어류치사성 Cochlodinium polykrikoides 적조생물의 유전적 진화 및 특성)

  • Cho, Eun-Seob;Jeong, Chang-Su
    • Journal of Life Science
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    • v.17 no.11
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    • pp.1453-1463
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    • 2007
  • This study presents a molecular phylogenetic analysis of the harmful dinoflagellate Cochlodinium polykrikoides, by use of partial sequence of small subunit (SSU) rRNA gene from most of the major taxa(24 species) in dinoflagellates. The class Dinophyceae clade formed a strong monophyletic relationship with C. polykrikoides and several taxa. On the basis of deeper nodes, the phylogenetic relationships placed C. polykrikoides closer to the order Prorocentrales rather than to the order Gymnodiniales, which was supported by a strong bootstrap value (100%) in the analyses of Neighbor-Joining and Parsimony methods. There is strong support for C. polykrikoides being placed in the same branch as Gymnodiniaceae and being connected in a clade with Prororcentrum micans among Prorocentrales. Morphological data show that C. polykrikoides is well associated with the genus Gyrodinium; however, this species is genetically closer to Gymnodinium than to Gyrodinium. The placement of C. polykrikoides always formed an independent branch separated from other dinoflagellates. In conclusion, planktonic P. micans plays an important role as an ancestor of Gymnodinium, whereas C. polykrikoides appears to be used an intermediate position between P. micans and Gymnodinium based on evolution.

Studies on Microbial Extracellular $\beta$-Gala-ctosidase

  • Lee, Keun-Eok
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 1979.04a
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    • pp.113.2-114
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    • 1979
  • $\beta-Galactosidase$ is an enzyme which catalizes hydrolysis of lactose, a natural substrate, to glucose and galctose and transferring some monosac-charide units to active acceptors as sugar or alcohol. The occurence of $\beta-Galactosidase$ is known in various microorganisms, animals and higher plants and has been studied by many investigatigators. Especially, a great deal of articles for the enzyme of E. coli have been presented in genetic control mechanism and induction-repression effects of proteins, On the other hand, in the dairly products industry, it is important to hydrolyes lactosd which is the principal sugar of milk and milk products. During the last few years, the interest in enzymatic hydrolysis of milk lactose has teen increased, because of the lactose intolerence in large groups of the population. Microbial $\beta-Galactosidases$ are considered potentially most suitable for processing milk to hydrolyse lactose and, in recent years, the immobilized enzyme from yeast has been examined. Howev, most of the microbial $\beta-Gal$ actosidase are intracellular enzymes, except a few fungal $\beta-Gala-$ ctosidases, and extracellular $\beta-Galactosidase$ which may be favorable to industrial applieation is not so well investigated. On this studies, a mold producing a potent extracellular $\beta-Galactosidase$ was isolated from soil and identified as an imperfect fungus, Beauveria bassians. In this strain, both extracellular and intracellular $\beta-Galactosidases$ were produced simultaneously and a great increase of the extracellular production was acheved by improving the cultural conditions. The extracellular enzyme was purified more than 1, 000 times by procedures including Phosphocellulose and Sephadex G-200 chromatographies. Several characteristics of the enzymewas clarified with this preparation. The enzyme has a main subunit of molecular weight of 80, 000 which makes an active aggregate. And at neutral pH range, it has optimum pH for activity and stability. The Km value was determined to be 0.45$\times$10$^{-3}$ M for $o-Nitrophenyl-\beta-Galactoside.$ In any event, it is interesting to sttudy the $\beta-Galactosidase$ of B. bassiana for the mechanism of secretion and conformational structure of enzyme.

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Morphological and Molecular Characterization of Desportesius invaginatus (Nematoda: Acuariidae) from Egretta garzetta and Bubulcus ibis in Korea

  • Lee, Seo-In;Hong, Eui-Ju;Kim, Hyeon-Cheol;Ryu, Si-Yun;Park, Bae-Keun
    • Journal of Veterinary Clinics
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    • v.38 no.2
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    • pp.75-81
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    • 2021
  • An aquariid nematode, Desportesius invaginatus, was found in the proventriculus of an Egretta garzetta and a Bubulcus ibis from Chuncheon in the Republic of Korea. The worms were identified by light and scanning electron microscopy based on important taxonomic characteristics (body length, esophagus length, cordons, spicules, caudal alae of males, position of the vulva) and then phylogenetically analyzed using the 18S rRNA encoding gene. The nematodes were characterized by a body length of 7.0-8.0 mm in males and 10.2-13.1 mm in females, and two pairs of cordons recurrent in the anterior direction, and cordons were anastomosed by a longitudinal cuticular ridge that externally delimits a longitudinal canal. The widest cuticular plates of cordons bears over 20 posterior spines. The length of the spicules in males was also significantly different. The right spicule measured 742-821 (794) ㎛ in length and 40-45 (42) ㎛ in width, and the left spicule measured 493-556 (541) ㎛ in length and 11-13 (12) ㎛ in width. The caudal alae of males are inflated and vesicular in appearance. The vulva was situated at 56-71 (58.3) ㎛ from the posterior extremity. Although the 18S rRNA sequences of worms were similar to the Synhimantus species, some genetic divergences were observed in comparison. In this study, the worms were recognized with genus Desportesius because genus Desportesius was considered a subgenus of Synhimantus. This is the first record of D. invaginatus in the Republic of Korea.

Clinical and molecular characterization of Korean children with infantile and late-onset Pompe disease: 10 years of experience with enzyme replacement therapy at a single center

  • Kim, Min-Sun;Song, Ari;Im, Minji;Huh, June;Kang, I-Seok;Song, Jinyoung;Yang, Aram;Kim, Jinsup;Kwon, Eun-Kyung;Choi, Eu-Jin;Han, Sun-Ju;Park, Hyung-Doo;Cho, Sung Yoon;Jin, Dong-Kyu
    • Clinical and Experimental Pediatrics
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    • v.62 no.6
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    • pp.224-234
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    • 2019
  • Purpose: Pompe disease (PD) is an autosomal recessive disorder caused by a deficiency of acid alpha-glucosidase resulting from pathogenic GAA variants. This study describes the clinical features, genotypes, changes before and after enzyme replacement therapy (ERT), and long-term outcomes in patients with infantile-onset PD (IOPD) and late-onset PD (LOPD) at a tertiary medical center. Methods: The medical records of 5 Korean patients (2 male, 3 female patients) diagnosed with PD between 2002 and 2013 at Samsung Medical Center in Seoul, Republic of Korea were retrospectively reviewed for data, including clinical and genetic characteristics at diagnosis and clinical course after ERT. Results: Common initial symptoms included hypotonia, cyanosis, and tachycardia in patients with IOPD and limb girdle weakness in patients with LOPD. Electrocardiography at diagnosis revealed hypertrophic cardiomyopathy in all patients with IOPD who showed a stable disease course during a median follow-up period of 10 years. Patients with LOPD showed improved hepatomegaly and liver transaminase level after ERT. Conclusion: As ERT is effective for treatment of PD, early identification of this disease is very important. Thus, patients with IOPD should be considered candidates for clinical trials of new drugs in the future.

Isolation and identification of mammalian orthoreovirus type 3 from a Korean roe deer (Capreolus pygargus)

  • Yang, Dong-Kun;An, Sungjun;Park, Yeseul;Yoo, Jae Young;Park, Yu-Ri;Park, Jungwon;Kim, Jong-Taek;Ahn, Sangjin;Hyun, Bang-Hun
    • Korean Journal of Veterinary Research
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    • v.61 no.2
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    • pp.13.1-13.8
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    • 2021
  • Mammalian reovirus (MRV) causes respiratory and intestinal disease in mammals. Although MRV isolates have been reported to circulate in several animals, there are no reports on Korean MRV isolates from wildlife. We investigated the biological and molecular characteristics of Korean MRV isolates based on the nucleotide sequence of the segment 1 gene. In total, 144 swabs from wild animals were prepared for virus isolation. Based on virus isolation with specific cytopathic effects, indirect fluorescence assays, electron microscopy, and reverse transcription-polymerase chain reaction, only one isolate was confirmed to be MRV from a Korean roe deer (Capreolus pygargus). The isolate exhibited a hemagglutination activity level of 16 units with pig erythrocytes and had a maximum viral titer of 105.7 50% tissue culture infectious dose (TCID50)/mL in Vero cells at 5 days after inoculation. The nucleotide and amino-acid sequences of the partial segment S1 of the MReo2045 isolate were determined and compared with those of other MRV strains. The MReo2045 isolate had nucleotide sequences similar to MRV-3 and was most similar (96.1%) to the T3/Bat/Germany/342/08 strain, which was isolated in Germany in 2008. The MReo2045 isolate will be useful as an antigen for sero-epidemiological studies and developing diagnostic tools.

Molecular Identification and Morphological Description of Larvae of the Previously Unrecorded Species Cryptacanthodes bergi (Zoarcoidei: Cryptacanthodidae) collected from Gangwon Province, Korea (강원도에서 채집된 등가시치아목 1미기록종, Cryptacanthodes bergi 자어의 분자동정 및 형태기재)

  • Choi, Si-Won;Lee, Soo-Jeong;Kim, Jin-Koo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.54 no.2
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    • pp.188-193
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    • 2021
  • On March 13, 2018 two postflexion larval specimens (18.28 mm and 16.80 mm in standard length) belonging to the family Cryptacanthodidae, suborder Zoarcoidei were collected from Sokcho and Gangneung in Gangwon Province. The family Cryptacanthodidae comprises 4 species worldwide: 3 in the North Pacific Ocean and 1 in the western North Atlantic Ocean. As a result of analyzing 620 bp of the mtDNA COI region, the two postflexion larvae collected in this study were identified as Cryptacanthodes bergi by 99.5% agreement with C. bergi adult registered in NCBI. Postflexion larvae of C. bergi are compressed with large eyes and radial pectoral fins and the anus located in front of the center of the body. Melanophores were intensively distributed along the dorsal midline, except for caudal peduncle, and sporadically distributed on the back of the anus. In addition, there were limited star-like melanophores on the back of the gut. This species showed 0.058 genetic distance when compared mtDNA COI region of C. aleutensis, and it was well distinguished in the distribution pattern of black vesicles of the head, count and measurement traits. Considering the morphological and ecological characteristics of this species, we suggest a new Korean name, " Gwisin-jang-gaeng-i ".

Modification of ginsenoside saponin composition via the CRISPR/Cas9-mediated knockout of protopanaxadiol 6-hydroxylase gene in Panax ginseng

  • Choi, Han Suk;Koo, Hyo Bin;Jeon, Sung Won;Han, Jung Yeon;Kim, Joung Sug;Jun, Kyong Mi;Choi, Yong Eui
    • Journal of Ginseng Research
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    • v.46 no.4
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    • pp.505-514
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    • 2022
  • Background: The roots of Panax ginseng contain two types of tetracyclic triterpenoid saponins, namely, protopanaxadiol (PPD)-type saponins and protopanaxatiol (PPT)-type saponins. In P. ginseng, the protopanaxadiol 6-hydroxylase (PPT synthase) enzyme catalyses protopanaxatriol (PPT) production from protopanaxadiol (PPD). In this study, we constructed homozygous mutant lines of ginseng by CRISPR/Cas9-mediated mutagenesis of the PPT synthase gene and obtained the mutant ginseng root lines having complete depletion of the PPT-type ginsenosides. Methods: Two sgRNAs (single guide RNAs) were designed for target mutations in the exon sequences of the two PPT synthase genes (both PPTa and PPTg sequences) with the CRISPR/Cas9 system. Transgenic ginseng roots were generated through Agrobacterium-mediated transformation. The mutant lines were screened by ginsenoside analysis and DNA sequencing. Result: Ginsenoside analysis revealed the complete depletion of PPT-type ginsenosides in three putative mutant lines (Cr4, Cr7, and Cr14). The reduction of PPT-type ginsenosides in mutant lines led to increased accumulation of PPD-type ginsenosides. The gene editing in the selected mutant lines was confirmed by targeted deep sequencing. Conclusion: We have established the genome editing protocol by CRISPR/Cas9 system in P. ginseng and demonstrated the mutated roots producing only PPD-type ginsenosides by depleting PPT-type ginsenosides. Because the pharmacological activity of PPD-group ginsenosides is significantly different from that of PPT-group ginsenosides, the new type of ginseng mutant producing only PPD-group ginsenosides may have new pharmacological characteristics compared to wild-type ginseng. This is the first report to generate target-induced mutations for the modification of saponin biosynthesis in Panax species using CRISPR-Cas9 system.