• Title/Summary/Keyword: PCR 동정

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Comparative Analysis of Genetic Variation of Cucumber Mosaic Virus from Commelina communis in Korea (국내 닭의장풀에서 분리된 오이모자이크바이러스 분리주들의 외피단백질 유전자와 병징 다양성 비교)

  • Park, Tae-Seon;Hong, Jin-Sung
    • Research in Plant Disease
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    • v.24 no.2
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    • pp.170-173
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    • 2018
  • Three isolates of Cucumber mosaic virus isolated from Commelina communis plants showing chlorosis and mosaic were collected in Chungju and Chuncheon, Korea. To confirm genetic variation of these three isolates (CMV-Co, CMV-Co2, and CMV-Co3), we performed PCR-RFLP and sequence analysis. Sequences of coat protein genes of CMV-C0, -Co2 and -Co3 were compared with CMV-Fny and showed 96.3%, 96.3%, and 95.9% similarities, respectively. In host reactions, three CMV-Co isolates induced systemic necrosis in Cucurbita pepo unlike CMV-Fny and CMV-Co, CMV-Co2 and CMV-Co3 observed differential symptoms responses in Physalis angulata and Nicotiana rustica. These results indicated that three isolates of CMV isolated from C. communis have genetic and biological variation.

Characterization of Rice black-streaked dwarf virus in Maize (옥수수에 발생하는 벼검은줄오갈병의 유전자 비교)

  • Lee, Bong-Choon;Yoon, Young-Nam;Hong, Sung-Jun;Hong, Yeon-Kyu;Hwang, Jae-Bok;Song, Sek-Bo;Kang, Hwang-Won;Lee, Key-Woon
    • Research in Plant Disease
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    • v.14 no.3
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    • pp.223-225
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    • 2008
  • This study was carried out to identify the Rice black-streaked dwarf virus that infected maize plants collected from Gochang-gun in Jeollabukdo in 2005. The genomic dsRNA from infected plants was extracted and the genome pattern was analyzed by polyacrylamide gel electrophoresis. Results of the electrophoresis revealed the already known to-segment genome and the difference of mobility was confirmed in isolates by collected areas. The RBSDV was identified from the result of RT-PCR using the template of extracted dsRNA and specific primer. The results of S10 cloned to pGEM-T vector and the conducted in sequence analysis consisted of 1,801nt and 559aa. This was of the same size as the RBSDV S10 identified in rice, and the change was confirmed in 18 base and displayed homology of 99%.

Isolation and Identification of Yeasts from Wild Flowers of Daejeon City and Chungcheongnam-do in Korea (대전과 충남 지역에서 채취한 야생화로부터 효모의 분리 및 동정)

  • Min, Jin-Hong;Hyun, Se-Hee;Kang, Min-Gu;Lee, Hyang-Beom;Kim, Chang-Mu;Kim, Ha-Kun;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.40 no.3
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    • pp.141-144
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    • 2012
  • Total 50 species, 80 strains: Eighty strains of yeasts were isolated from wild flowers of Daejeon city and its surroundings in Korea. Isolated yeasts were identified by comparison of their PCR-amplified 18s rDNA V3 region or internal transcribed spacer-2 (ITS-2) genes with BLAST database. Among them, 32 species, 54 strains of yeasts were isolated and identified from wild flowers of Daejeon city, and Cryptococcus species were dominant yeasts. Furthermore, 18 species, 26 strains of yeasts were isolated and identified from wild flowers of Geumsan-gun and Nonsan-gun, Chungcheongnam-do in Korea, and also Candida species and Pseudozyma species were dominant yeasts.

Increased Expression of aac(3)II by Tn3 in Gentamicin - Resistant Bacteria Isolated from Hospital Sewage (병원하수로부터 분리한 Gentamicin 저항성 세균에서 Tn3에 의한 aac(3)II의 발현 증가)

  • 한효심;이문숙;정재성
    • Korean Journal of Microbiology
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    • v.40 no.1
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    • pp.60-64
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    • 2004
  • We tested gentamicin - resistant bacteria isolated from hospital sewage to confirm the presence of aac(3)II encoding aminoglycoside- (3)-N- acetyltransferase by dot-blot hybridization. A probe from the internal fragment of aac(3)II was hybridized to DNA from 41 % (39/95) of gentamicin resistant isolates. PCR was performed with primers from aac(3)II and Tn3. Of 39 strains, 13 strains had Tn3-aac(3)II structure. Minimal inhibitory concentration (MIC) test demonstrated that 18 strains containing Tn3-aac(3)II showed higher resistance to gentamicin than those of other strains. Thirteen strains were identified as 5 Escherichia coli, 3 Acinetobacter johnsonii, 2 Enterobacter agglomerans, 2 Micrococcus luteus, and 1 Pseudomonas facilis. These results suggest that gentamicin-resistant determinant of Tn3-aac(3)II structure was widely distributed in the gentamicin-resistant bacteria.

A study on the Aptamer Specific Detection on P. gingivalis (P. gingivalis에 특이적으로 작용하는 앱타머에 관한 연구)

  • Shin, Ae-Ri
    • The Journal of the Korea Contents Association
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    • v.21 no.4
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    • pp.825-832
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    • 2021
  • In this study, by selecting specific aptamers that selectively detection on P. gingivalis, the main cause of periodontal disease, and purifying and identifying protein molecules that bind to the selected aptamers, the mechanism of action of P. gingivalis was investigated. A DNA library having 39 random sequences was prepared, and aptamers with specificity for P. gingivalis were selected using the SELEX method, and the nucleotide sequence was analyzed by cloning using PCR2.1 cloning vector. 8 of aptamers with different nucleotide sequences were selected, and modified weston blot was performed using APG-3 among the selected aptamers to identify 11 proteins that act directly, and proteins were analyzed. As a result, a protein that selectively binds to P. gingivalis was isolated and identified. Therefore, aptamer selectively binds and attaches to proteins related to inhibition of sugar metabolism and cell activity of P. gingivalis, suggesting the possibility of a sensor for diagnosis of periodontal disease.

Isolation and identification of culturable bacteria from human skin (배양가능한 피부세균의 분리 및 동정)

  • Bae, Young-Min
    • Journal of the Korean Applied Science and Technology
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    • v.37 no.6
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    • pp.1698-1705
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    • 2020
  • Bacteria were collected from the thumb surface of the twenty young adults that are 20 to 25 years old and cultured on the Luria-Bertani agar. The 16S rDNA of the cultured bacteria was amplified by polymerase chain reaction(PCR) and DNA sequence of the PCR products analyzed. Total 14 different bacterial species were identified by comparing their 16S rDNA sequence with the data in genbank. It appears that each individual has 2.5 different bacterial species in average. Staphylococcal species were the most abundant among the identified bacteria and Micrococcus luteus was the second. Staphylococcal species were isolated at similar frequency between male and female donors but Micrococcus luteus was isolated more frequently from female than male donors. The result obtained in this study might be useful in research of dermatic diseases, searching for new drugs for those diseases and development of new cosmetics.

A study on the characteristics and pathogenicity of Aeromonas veronii isolated from infected goldfish (Carassius auratus) (피부 궤양이 발생한 금붕어(Carassius auratus)에서 분리한 Aeromonas veronii의 특성 및 병원성 분석)

  • Hyeon Ki Jung;Min Su Kim;Sok Ho Kim;Min Soon Choi
    • Journal of fish pathology
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    • v.37 no.1
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    • pp.79-88
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    • 2024
  • Aeromonas spp. infections have been reported to cause significant economic losses not only in the ornamental fish industry but also in aquaculture. In December 2022-January 2023, an Aeromonas infection occurred on a goldfish in korea, A gram-negative bacterium was isolated from the skin and internal organs of infected goldfish (Carassius auratus). The results showed that the isolate was identified as Aeromonas veronii using 16S rDNA targeted oilgpnucleotide primers, furthermore characteristics of A. veronii was confirmed by enterotoxin gene, infectious experiment, antibiotic resistance. In-vivo pathogenicity of isolates to goldfsh resulted in 100% mortality in challenged host within one week of post experiment injection. As a result of PCR analysis targeting three enterotoxin-encoding genes, cytotoxic enterotoxin (act) was identified in A. veronii isolate in this study. Antimicrobial susceptibility pattern of isolate showed it was to susceptible to most antimicrobial agents tested but resistant to ampicillin, imipenem, meropenem and clindamycin.

Identification of Korean Ginseng Cytochrome P450 gene and Its Characterization by Transformation System (고려인삼 유래 Cytochrome P450 유전자의 동정 및 형질전환에 의한 특성검정)

  • Shim, Ju-Sun;Kim, Yu-Jin;Jung, Seok-Kyu;Kwon, Woo-Saeng;Kim, Se-Young;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • v.33 no.3
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    • pp.212-218
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    • 2009
  • Triterpenoid saponins were synthesized in Panax ginseng C.A. Meyer via the isoprenoid pathway by cyclization of 2,3-oxidosqualene to give primarily oleanane (beta-amyrin) or dammarane triterpenoid skeletons. The triterpenoids are backbone and undergoes various modifications (oxidation, substitution and glycosylation), mediated by cytochrome P450 (CYP)-dependent monooxygenases, glycosyltransferase and other enzymes. This is likely to be due in part to the complexity of the molecules and the lack of pathway intermediates for biochemical studies. A cDNA clone encoding a putative CYP gene was isolated from flower bud of ginseng and transformed into the plant(Nicotiana tabacum cv. Xanthi) and confirmed by PCR analysis. The CYP gene (PgCYP) contained an open reading frame(ORF) encoding mature protein of 500 amino acids. The expression of PgCYP were investigated in transgenic tobacco by reverse transcriptase-polymerase chain reaction (RT-PCR).

Cloning and Identification of Differentially Expressed Genes Induced by Fungal Infection from Silkworm, Bombyx mori (누에에서 곰팡이(Aspergillus niger) 감염에 의해 유도 발현되는 유전자의 클로닝과 동정)

  • Lee, Jin-Sung;Hong, Su-Young;Lee, Ki-Hwa
    • Journal of Life Science
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    • v.20 no.6
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    • pp.929-933
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    • 2010
  • We tried to identify differentially expressed genes (DEGs) from a silkworm, Bombyx mori, involved in fungal (Aspergillus niger) infection. A total RNA purified from fungal-induced and normal B. mori ($5^{th}$ instar larvae) was used for the cDNA synthesis. Differentially expressed genes were screened by annealing control primer (ACP)-based PCR technique. Comparing the gene expression profiles between fungal infection and control silkworm, we detected 10 genes that were differentially expressed in fungal induction and performed molecular cloning and nucleotide sequencing of the 10 genes. We confirmed the expression patterns of 3 DEGs by RT-PCR. The 3 DEGs over-expressed in fungal infection were identified as lysozyme, enbocin and an unknown gene. They were first identified to be genes induced by fungal infection. Although the detailed functions of 3 genes and their products remain to be determined, the genes will provide insight into the molecular mechanisms of insect-immune systems induced by fungal infection.

Identification of Ruditapes philippinarum and Meretrix lusoria Larvae Using Single Cell PCR Analysis and Microscopic Observation (Single Cell PCR과 현미경을 통한 바지락 및 백합 유생의 동정)

  • Jung, Seung-Won;Kim, Chang-Soo;Yoo, Jae-Won;Kim, Young-Ok;Lee, Jin-Hwan;Hong, Jae-Sang
    • Ocean and Polar Research
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    • v.32 no.3
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    • pp.247-254
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    • 2010
  • Single cell PCR analysis and light and scanning electron microscopic techniques were utilized to identify free living bivalve larvae in the coastal waters of Tae-an, on the west coast of Korea. Through DNA sequencing, venerid clam larvae were isolated and identified as Ruditapes philippinarum (99% similarity) and Meretrix lusoria (99%). Under microscopic observation, the D-veliger stage of R. philippinarum exhibited symmetrical shoulder angles and an elliptical ventral form. In contrast, M. lusoria displayed asymmetrical shoulder angles and a round ventral form in the umbonal stage. Size of the R. philippinarum larvae was $156{\pm}22{\mu}m$ in length, $126{\pm}12{\mu}m$ in height, $92{\pm}14{\mu}m$ in width with a length: height ratio of 1.23. Meretrix lusoria was $202{\pm}44{\mu}m$ in length, $161{\pm}35{\mu}m$ in height, $96{\pm}38{\mu}m$ in width with a length: height ratio of 1.25. Experimental results indicate that morphological and molecular characteristics provide evidence for the larval identification of these two venerid clam larvae species in nature.