• Title/Summary/Keyword: Nucleotide sequence

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Epidemiological surveys of an outbreak of false positive classical swine fever in Gyeongbuk province (의사 돼지콜레라 발생농장 역학조사)

  • Park, No-Chan;Cho, Kwang-Hyun;Kim, Young-Hoan;Kim, Soon-Tae;Kim, Sung-Kuk;Park, In-Hwa;Cho, Min-Hee;Oh, Gang-Hee;Son, Jae-Kweon;Jyeong, Jong-Sik
    • Korean Journal of Veterinary Service
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    • v.26 no.2
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    • pp.163-184
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    • 2003
  • This study was conducted to survey the farm which suffered from disease similar to classical swine fever(CSF) in Gyeongbuk province. Clinical signs appeared first in a few number of growing pigs which showed specific signs of diarrhea, depression, sleepiness, and reluctance to get up or to eat. Younger piglets may have appeared chilled, shiver and huddle together. As the disease progresses the affected pig's skin went red and purple. In histopathological signs, there were many haemorrhages throughout the body and larger haemorrhages in some organs such as lymph nodes. And there is a precipitous fall in the number of circulating leukocytes in the blood. In spite of insisting of farmer which did not vaccinate to classical swine fever, significant antibody production was detected in these affected pigs at enzyme-linked immuonsorbent assay. According to the above results at first glance, these affected pig suspected with CSF in clinical signs and histopathological lesions only. Because the symptoms and post-mortem picture were very similar to CSF, these false positive results would have been dangerous to diagnostician. But by reverse transcriptase polymerase chain reaction(RT-PCR) and comparative nucleotide sequence analysis, the disease was correctly diagnosed with post-weaning multisystemic wasting syndrome(PMWS) and porcine reproductive and respiratory syndrome(PRRS) compoundly. And the antigen which were detected the lesion similar to CSF virus, was confirmed with LOM vaccine strain of CSF. In most national CSF eradication program and in countries which are free of the CSF virus, vaccination against CSF is not practiced and generally is not allowed. But now in Korea, routine vaccination is practiced because of outbreaking the CSF repeatedly. When CSF is diagnosed the whole herd and other in contact animal are slaughtered continuously.

Isolation of indigenous Lactobacillus plantarum for malolactic fermentation (말로락틱 발효에 적합한 토착 Lactobacillus plantarum 분리)

  • Heo, Jun;Lee, Chan-Mi;Park, Moon Kook;Jeong, Do-Youn;Uhm, Tai-Boong
    • Korean Journal of Microbiology
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    • v.51 no.2
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    • pp.169-176
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    • 2015
  • The malolactic fermentation (MLF), which is widely used in winemaking, is the conversion of malic acid to lactic acid conducted by the malolactic enzyme (Mle) of lactic acid bacteria. In order to select the strains with MLF among 54 lactic acid bacteria isolated from the traditionally fermented foods, we designed a primer set that specifically targets the conserved regions of the mle gene and then selected four strains that harbor the mle gene of Lactobacillus plantarum. All strains were identified as L. plantarum by analyzing the 16S rRNA sequences, biochemical properties, and the PCR products of the recA gene. From comparison of the mle gene sequences consisting of 1,644 bp, the nucleotide and amino acid sequence of strain JBE60 correspond to 96.7% and 99.5% with those of other three strains, respectively. The strain JBE60 showed the highest resistant against 10% (v/v) ethanol among the strains. The strains lowered the concentration of malic acid to average 43%. Considering the ethanol resistance and conversion of malic acid, the strain JBE60 is considered as a potential starter for the malolactic fermentation.

In vitro activity comparison of Erm proteins from Firmicutes and Actinobacteria (Firmicutes와 Actinobacteria에 속하는 세균들의 Erm 단백질 in vitro 활성 비교)

  • Jin, Hyung Jong
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.269-277
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    • 2016
  • Erm proteins methylate the specific adenine residue ($A_{2058}$, E. coli numbering) on 23S rRNA to confer the $MLS_B$ (macrolidelincosamide-streptogramin B) antibiotic resistance on a variety of microorganisms ranging from antibiotic producers to pathogens. When phylogenetic tree is constructed, two main clusters come out forming each cluster of Actinobacteria and Firmicutes. Two representative Erm proteins from each cluster were selected and their in vitro methylation activities were compared. ErmS and ErmE from Actinobacteria cluster exhibited much higher activities than ErmB and ErmC' from Firmicutes: 9 fold difference when ErmC' and ErmE were compared and 13 fold between ErmS and ErmB. Most of the difference was observed and presumed to be caused by N-terminal and C-terminal extra region from ErmS and ErmE, respectively because NT59TE in which N-terminal end 59 amino acids was truncated from wild type ErmS exhibited only 22.5% of wild type ErmS activity. Meanwhile, even NT59TE showed three and 2.2 times more activity when it was compared to ErmB and C, respectively, suggesting core region from antibiotic producers contains extra structure enabling higher activity. This is suggested to be possible through the extra region of 197RWS199 (from both ErmS and ErmE), 261GVGGSLY267 (from ErmS), and 261GVGGNIQ267 (from ErmE) and 291SVV293 (from ErmS) and 291GAV293 (from ErmE) by multiple sequence alignment.

Bioequivalence of Pinatos Capsule 10 mg to Ketas Capsule 10 mg (Ibudilast 10 mg) (케타스 캡슐 10밀리그램(이부딜라스트 10 밀리그램)에 대한 피나토스 캡슐 10밀리그램의 생물학적동등성)

  • Kang, Hyun-Ah;Kim, Se-Mi;Kang, Min-Sun;Yoo, Dong-Jin;Lee, Sang-No;Kwon, In-Ho;Yoo, Hee-Doo;Lee, Yong-Bok
    • Journal of Pharmaceutical Investigation
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    • v.40 no.2
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    • pp.117-123
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    • 2010
  • Ibudilast, 3-isobutyryl-2-isopropyrazolo[1,5-a]pyridine, is a nonselective inhibitor of cyclic nucleotide phosphodiesterase (PDE). It preferentially inhibits PDE 3A, PDE4, PDE10 and PDE11 as well as a number of the other PDE families, albeit to a lesser extent. Ibudilast is used clinically to treat bronchial asthma and cerebrovascular disorders. Thes e clinical uses are based on the ability of ibudilast to inhibit platelet aggregation, improve cerebral blood flow and attenuate allergic reactions. The purpose of the present study was to evaluate the bioequivalence of two ibudilast capsules, Ketas capsule (Handok Pharmaceuticals Co., Ltd.) and Pinatos capsule (Sam Chun Dang Pharm. Co., Ltd.), according to the guidelines of the Korea Food and Drug Administration (KFDA). The in vitro release of ibudilast from the two ibudilast formulations was tested using KP Apparatus method with various dissolution media. Twenty six healthy male subjects, 23.31${\pm}$1.09 years in age and 70.45${\pm}$8.51 kg in body weight, were divided into two groups and a randomized $2{\times}2$ cross-over study was employed. After a single capsule containing 10 mg as ibudilast was orally administered, blood samples were taken at predetermined time intervals and the concentrations of ibudilast in serum were determined using HPLC/UV detector. The dissolution profiles of two formulations were similar in all tested dissolution media. The pharmacokinetic parameters such as $AUC_t$, $C_{max}$ and $T_{max}$ were calculated, and computer programs (Equiv Test and K-BE Test 2002) were utilized for the statistical analysis of the parameters using logarithmically transformed $AUC_t$, $C_{max}$ and untransformed $T_{max}$. The results showed that the differences between two formulations based on the reference drug, Ketas, were 6.99%, -2.48% and 9.93% for $AUC_t$, $C_{max}$ and $T_{max}$, respectively. There were no sequence effects between two formulations in these parameters. The 90% confidence intervals using logarithmically transformed data were within the acceptance range of log 0.8 to log 1.25 (e.g., log 0.8791~log 1.1861 and log 0.8347~log 1.1199 for $AUC_t$ and $C_{max}$, respectively). Thus, the criteria of the KFDA bioequivalence guideline were satisfied, indicating Pinatos capsule was bioequivalent to Ketas capsule.

Distinguishing the Korean Silage Corn Varieties through Development of PCR-Based SNP Marker (SNP마커 개발을 통한 사료용 옥수수 품종판별)

  • Kim, Sang Gon;Lee, Jin-Seok;Bae, Hwan Hee;Kim, Jung-Tae;Son, Beom-Young;Baek, Seong-Bum
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.37 no.2
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    • pp.168-175
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    • 2017
  • Single nucleotide polymorphisms (SNP) markers allow rapid screening of crop varieties in early growth stages. We developed a modified SNP PCR procedure for assaying SNPs in maize. For SNP marker development, we chosen 200 SNP sites from MaizeGDB database, and designed two base pair mismatch primers based on putative SNP site of B73 genome sequence. PCR products size was from 200 to 500 bp or was not shown in the case of SNP site existing in Korean silage corns. Using previously discovered 16 primer sets, we investigated distinctness of 50 silage F1 hybrid corns including 10 Korean silage corns developed by RDA such as Gangdaok, Kwangpyeongok, Dapyeongok, Andaok, Yanganok, Singwangok, Jangdaok, Cheongdaok, Pyeonggangok, and Pyeonganok as well as 40 foreign commercial silage corns. From cluster analysis, we confirmed that 10 Korean silage F1 hybrid corns were clearly distinguished except for Singwangok, P1395, and several foreign commercial corns, and selected minimum SNP primer combination for Gangdaok, Jangdaok, Pyeonggangok, and Pyeonganok. Therefore, development of SNP marker sets might be faster, cheaper, and feasible breed discrimination method through simple PCR and agarose gel electrophoresis.

Novel Variations in Human Interleukin-29 and Their Association (사람의 Interleukin-29 유전자의 새로운 변이의 단리 및 그들의 연관)

  • Song, Ju-Hee;Chae, Soo-Cheon;Lee, Jae-Hoon;Chung, Hun-Taeg
    • Journal of Life Science
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    • v.14 no.2
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    • pp.275-279
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    • 2004
  • Gene polymorphisms of cytokines and their receptors are attractive candidates as genetic factors in the pathogenesis of immune-mediated diseases and have been reported to be associated with disease susceptibility to autoimmune, inflammatory and infectious diseases. IL-29 is one of important candidate genes for complex trait of genetic diseases but there is no published survey of single nucleotide polymorphisms (SNPs) in this gene. In this study, for the first time, we have examined the full genomic sequence of IL-29 including the promoter regions to identify SNPs. We examined the frequencies of genotypes and alleles at the SNP site of IL-29 in allergic rhinitis patients and non-allergic rhinitis controls using the direct sequencing method to determine whether this IL-29 SNP is associated with allergic rhinitis in Korean population. We identified one novel SNP (1184C>A) in the intron 2 and one novel variation site (-1842_-1841dupGA) in the promoter region of human IL-29 gene. The P values of SNP or variation site were not significant between the healthy controls and allergic rhinitis patients. Our results suggest that the 1184C>A polymorphism and -1842_-1841dupGA variation site in human IL-29 gene were not associated to allergic rhinitis.

Diversity and Plant Growth-Promotion of Endophytic Fungi Isolated from the Roots of Plants in Dokdo Islands (독도의 자생식물 뿌리에서 분리한 내생진균의 다양성과 생장촉진활성)

  • You, Young-Hyun;Yoon, Hyeok-Jun;Lee, Gil-Seong;Woo, Ju-Ri;Rim, Soon-Ok;Shin, Jae-Ho;Lee, In-Jung;Choo, Yeon-Sik;Kim, Jong-Guk
    • Journal of Life Science
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    • v.21 no.7
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    • pp.992-996
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    • 2011
  • Endophytic fungi were isolated from the roots of plants growing naturally on the island of Dokdo. Plant samples, such as Miscanthus sinensis, Achyranthus japonica and Echinochloa crusgali were isolated from Dongdo, and those such as Honkenya peploides and Artemsia koidzumii were isolated from Seodo. Twenty one strains of endophytic fungi were isolated from these plants. To identify the strains, PCR (polymerase chain reaction) amplification of the partial ITS (Internal Transcribed Spacer) regions was done with universal primers ITS-1 and ITS-4 to determine the nucleotide sequence of the ITS regions. Of the strains isolated from Miscanthus sinensis, 75% were Penicillium sp. and 25% were Aspergillus sp. Fifty five percent of strains isolated from Achyranthus japonica were Penicillium sp., 30% were Aspergillus sp. and 15% were Zygorhynchus sp. Strains isolated from Echinochloa crusgali were Penicillium sp. (50%), Aspergillus sp. (12%), Giberella sp. (13%), Talaromyces sp. (9%) and Umbelopsis sp. (8%). Of the strains isolated from Honkenya peploides, 76% were Penicillium sp. and 24% were Pestalotiopsis sp. Strains isolated from Artemisia koidzumii were Penicillium sp. (81%) and Mucor sp. (19%). As a result of bioassay, Ec-3-1 strain isolated from Echinochloa crusgalli showed plant growth-promotion activity. Of all the endophytic fungi isolated, Penicillium sp. was the most abundantly distributed fungal strain in all plants used in this study.

Phylogenetic Analysis of Korean Black Cattle Based on the Mitochondrial Cytochrome b Gene (mtDNA cytochrome b에 기초한 한국흑우의 계통유전학적 분석)

  • Kim, Jae-Hwan;Byun, Mi Jung;Kim, Myung-Jick;Suh, Sang Won;Kim, Young-Sin;Ko, Yeoung-Gyu;Kim, Sung Woo;Jung, Kyoung-Sub;Kim, Dong-Hun;Choi, Seong-Bok
    • Journal of Life Science
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    • v.23 no.1
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    • pp.24-30
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    • 2013
  • The purpose of this study was to identify genetic polymorphisms of the mitochondrial cytochrome b (mtDNA cyt b) gene in Korean black (KB) cattle breed and to analyze the genetic relationship between the KB and other breeds. We determined the complete sequence of the mtDNA cyt b gene in 38 KB cattle. We also analyzed their genetic diversity, and phylogenetic analysis was performed by comparison with Korean cattle (KC, called Hanwoo) and breeds from China and Japan. A nucleotide substitution was detected in the KB cattle, and two haplotypes were defined. In the neighbor-joining (NJ) tree, the haplotypes of KB were located in Bos taurus lineage with those of KC, Japanese black (JB), Yanbian and Zaosheng breeds. However, the haplotypes of Chinese breeds, excluding Yanbian and Zaosheng, were separated into B. taurus and B. indicus lineages. In the NJ tree of breeds based on Dxy genetic distances, Chinese breeds mixed with B. taurus and B. indicus lineages were located between B. indicus and B. taurus lineages. KB was contained within B. taurus lineage and was determined to be genetically more closely related to two Chinese (Yanbian and Zaosheng) breeds than to KC and JB. The haplotype distribution and the results of the phylogenetic analysis suggest that KB and KC have genetic differences in their mtDNA cyt b gene sequences.

Purification and Characterization of the Bacteriocin Produced by Lactococcus sp. KD 28 Isolated from Kimchi (김치에서 분리한 Lactococcus lactis가 생산하는 박테리오신의 정제 및 특성)

  • Lee, Ji-Young;Choi, Nack-Shick;Chun, Sung-Sik;Moon, Ja-Young;Kang, Dae-Ook
    • Journal of Life Science
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    • v.25 no.2
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    • pp.180-188
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    • 2015
  • The bacterial strain isolated from Kimchi showed antibacterial activity against Micrococcus luteus IAM 1056. The selected strain was identified as Lactococcus lactis by 16S rRNA nucleotide sequence analysis and named as Lactococcus sp. KD 28. The treatment of culture supernatant with proteinase K removed antibacterial activity, indicating its proteinaceous nature, a bacteriocin. This bacteriocin was sensitive to hydrolytic enzymes such as ${\alpha}$-chymotrypsion, trypsin, proteinase K, lipase, ${\alpha}$-amylase and subtilisin A. The bacteriocin was highly thermostable and resistant to heating at $80^{\circ}C$ for up to an hour but 50 % of the total activity was remained at $100^{\circ}C$ for 30 min. The pH range from 2.0 to 8.0 had no effect on bacteriocin activity and it was not affected by solvents such as acetonitrile, isopropanol, methanol, chloroform and acetone up to 50% concentration. The bacteriocin showed antibacterial activity against M. luteus IAM 1056, Lactobacillus delbrueckii subsp. lactis KCTC 1058, Enterococcus faecium KCTC 3095, Bacillus cereus KCTC 1013, B. subtilis KCTC 1023, Listeria ivanovii subsp. ivanovii KCTC 3444, Staphylococcus aureus subsp. aureus KCTC 1916, B. megaterium KCTC 1098 and B. sphaericus KCTC 1184. The bacteriocin was purified through ammonium sulfate concentration, SP-Sepharose chromatography and RP-HPLC. The molecular weight was estimated to be about 3.4 kDa by tricine-SDS-PAGE analysis.

Development of a Recombinant Streptomyces griseus with sprA and sprB Genes for Proteolytic Enzyme Production (Streptomyces griseus IFO13350 유래 sprA 및 sprB 유전자를 이용한 Pretense 생산균주 개발)

  • Hwang Ji-Hwan;Lee Chang-Kwon;Lee Kang-Mu;Jo Byoung-Kee;Park Hae-Ryong;Hwang Yong-Il
    • Korean Journal of Microbiology
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    • v.41 no.1
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    • pp.87-92
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    • 2005
  • Pronase, a protease produced for commercial purpose by Streptomyces griseus, was composed of serine protease, alkaline protease, aminopeptidase and carboxypeptidase complex, and it has been widely used as anti-inflammatory drugs for human therapy. In this study, we developed a new integration vector, pHJ101 derived from pSET152, containing strong promoter, ermE, to overexpress a certain protease gene. Specific PCR primers for cloning of sprA (a gene for S. griseus protease A) and sprB (a gene for S. griseus protease B) genes were designed from the basis of nucleotide sequence in databases and amplified by PCR. Plasmid pHJ201 and pHJ202 were constructed by inserting of amplified each gene in a vector pHJ101. S. griseus HA and S. griseus HB were respectively obtained by conjugal process of a parent strain, S. griseus IFO 13350 with the recombinant Escherichia coli harboring plasmid pHJ201 or pHJ202. When protease activity was measured in flask cultivation, produced protease levels of S. griseus HA and S. griseus HB increased about 5.3 times and 5 times, respectively, more than that of parent strain. And, the constructed integrating plasmid pHJ101 was applicable for overexpression of a certain gene in Streptomyces sp.