• Title/Summary/Keyword: PCR 동정

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Prevalence of Anaplasma sp. in Thrushes (Family Turdidae) in Jeju Island, Republic of Korea (제주도와 마라도내 지빠귀과 조류에서 Anaplasma spp. 감염 조사)

  • Oh, Mi-Rae;Moon, Kyoung-Ha;Kim, So-Yeon;Kim, Yun-Gi;Choi, Chang-Yong;Kang, Chang-Wan;Kim, Hwa-Jung;Lee, Kyoung-Kap;Yun, Young-Min
    • Journal of Veterinary Clinics
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    • v.31 no.3
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    • pp.206-211
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    • 2014
  • Anaplasmosis is a rickettsial zoonosis mediated by blood-sucking arthropods, such as ticks, flies, and mosquitos. Migratory birds are common hosts of ticks that are mediators of anaplasmosis, in particular, the tick infection rate in thrushes (family Turdidae) has been known to be high. The main purpose of this study is to survey the occurrence and prevalence of Anaplasma spp. from the migratory thrushes in Jeju island. We collected blood samples from 6 thrushes rescued at the Jeju Wildlife Rescue Center and from 34 wild-caught thrushes on Mara island which is a satellite island of Jeju. As a result, the nested PCR confirmed that seven out of 40 individuals (17.5%) were infected by Anaplasma spp. and all of them were identified as A. phagocytophilum based on sequences obtained from partial 16S rRNA. All the infected birds were on their northward migration in spring, our results suggest that the Turdidae family, which is a common and abundant migrant group passing through Jeju island, may act a role as active reservoir and disperser of A. phagocytophilum causing potential influx of the zoonotic pathogens from its wintering grounds in lower latitude to the mainland Korea as well as Jeju.

Identification and sequence analysis of small subunit ribosomal RNA gene of bovine Theileria isolates from Korea and Japan (한국과 일본 소에 감염된 Theileria 분리주의 small subunit ribosomal 유전자의 동정 및 분석)

  • Chae, Joon-seok;Park, Jin-ho;Kwon, Oh-deog;Waghela, Suryakant D.;Holman, Patricia J.;Wagner, Gerald G.;Lee, Joo-mook
    • Korean Journal of Veterinary Research
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    • v.38 no.4
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    • pp.909-917
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    • 1998
  • Six different sequences types(A through E and H) and a subtype(Bl) of the small subunit ribosomal RNA(SSUrRNA) gene were found in bovine Theileria isolates from different areas of Korea and Japan. The sequences were aligned and three hypervariable regions were observed in the nucleotide position ranges 212~231, 261~270 and 632~690. Five of the Theileria isolates yielded sequence type A; these were the field isolates KCB, KCN, and KCJ, and the laboratory stock KLS, all from Korea, and a single isolate from Japan (JHS). This sequence type is identical to the SSUrRNA gene sequence listed for Theileria buffeli (GenBank Accession No. Z15106) from Marula, Kenya. The Korean field isolate KKB yielded only a single sequence type (B), but multiple sequence types were found in some isolates. For example, KCB and JHS isolates yielded both types A and B ; isolate KKW showed types B and H; isolate KCN showed types A, C, and D ; and isolate KCJ showed types A, B, E, and a subtype B1. Finding of the multiple sequences SSUrRNA gene sequences suggests that bovine Theileria isolates from both Korea and Japan may consist of mixed populations.

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Comparison of the Effects of Gamma Ray and Electron Beam Irradiation to Improve Safety of Spices for Meat Processing (육가공용 향신료의 위생화를 위한 감마선 및 전자선 조사 효과 비교)

  • Kim, Byung-Hee;Kim, Hyun-Joo;Yoon, Yo-Han;Shin, Myung-Gon;Lee, Ju-Woon
    • Food Science of Animal Resources
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    • v.30 no.1
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    • pp.124-132
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    • 2010
  • This study evaluated the effects of gamma ray and electron beam (E-beam) to improve the safety of spices for meat processing. The spices (garlic powder, curry powder, turmeric powder, black pepper, white pepper, oregano, parsley, laurel leaf powder, basil, and rosemary) were irradiated by gamma ray and E-beam at 0, 2, 4, 6, 8, and 10 kGy. Total bacterial populations were then enumerated on total plate count agar, and bacteria isolated from the samples were identified by polymerase chain reaction (PCR). In addition, $D_{10}$ values for Bacillus cereus and Staphylococcus aureus inoculated in spices was determined, and the Ames test was conducted for genotoxicity analysis. The contaminated total bacterial populations in spices ranged from 1.5 to 3.8 Log CFU/g, and most of identified bacteria were Bacillus spp., and Staphylococcus spp. However, the bacterial populations decreased below the detection limit (2 Log CFU/g) after irradiation at 4 kGy except for parsley, which required 6 kGy in gamma ray and 8 kGy in E-beam to decrease total bacterial populations below detection limit. $D_{10}$ values were also higher (p<0.05) in E-beam treated samples than gamma-ray treated samples. No genotoxicity was observed in both conditions with and without metabolic activation. These results indicate that gamma ray (>4 kGy and <6 kGy) could be more useful to improve food safety of meat processing spices compared to E-beam.

Antimicrobial Effects of Retort and Gamma Irradiation on Bacterial Populations in Spicy Chicken Sauce (레토르트 및 감마선 조사에 의한 화닭 덮밥 소스의 미생물 제어 효과 비교)

  • Kim, Young-Sik;Kim, Hyun-Joo;Yoon, Yo-Han;Shin, Myung-Gon;Kim, Cheon-Jei;Shin, Mee-Hye;Lee, Ju-Woon
    • Food Science of Animal Resources
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    • v.30 no.1
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    • pp.141-147
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    • 2010
  • This study evaluated the antimicrobial effects of retort process and gamma irradiation on reduction of total bacterial populations in spicy chicken sauce, which is served on top of the steamed rice. Commercial spicy chicken sauce was treated with retort and gamma ray at 0, 1, 3, 5, and 10 kGy. Total aerobic bacterial populations were then enumerated on plate count agar and isolated bacteria from the test samples were identified using PCR analysis. Moreover, gamma ray sensitivity of identified bacteria was evaluated by $D_{10}$ values, and genotoxicity of gamma-irradiated samples was examined. Gamma irradiation at 3 kGy reduced total aerobic bacterial cell counts in spicy chicken sauce below detection limit, but total aerobic bacterial cell counts in test samples treated with retort were 2.1 log CFU/g. Identified bacteria from the samples were Bacillus subtilis, B. amyloiquefaciense, and B. pumils, and the $D_{10}$ values for B. subtilis and B. cereus were 0.39 ($R^2\;=\;0.921$) and 0.28 log CFU/g ($R^2\;=\;0.904$), respectively. The SOS chromotest showed that the gamma-irradiated spicy chicken sauce did not cause mutagenicity. These results indicate that gamma irradiation of spicy chicken sauce could be useful in ensuring microbial safety.

Gene Cloning and Expression of Trehalose Synthase from Thermus thermophilus HJ6 (Thermus thermophilus HJ6 유래 내열성 Trehalose Synthase의 유전자 클로닝 및 발현)

  • Kim, Hyun-Jung;Kim, Han-Woo;Jeon, Sung-Jong
    • Microbiology and Biotechnology Letters
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    • v.36 no.3
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    • pp.182-188
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    • 2008
  • A hyperthermophilic bacteria (strain HJ6) was isolated from a hot springs located in the Arima-cho, Hyogo, Japan. The cells were long-rod type ($2-4{\mu}m$), about $0.4{\mu}m$ in diameter. The pH and temperature for optimal growth were 6.5 and $80^{\circ}C$, respectively. Phylogenetic analysis based on the 16S rDNA sequence and biochemical studies indicated that HJ6 belonged to the genus Thermus thermophilus (Tt). The gene encoding the Trehalose synthase (TS) was cloned and sequenced. The open reading frame (ORF) of the TtTS gene was composed of 2,898 nucleotides and encoded a protein (975 amino acids) with a predicted molecular weight of 110.56 kDa. The deduced amino acid sequence of TtTS showed 99% and 83% identities to the Thermus caldophilus TS and Meiothermus ruber TS, respectively. TtTS gene was expressed in Escherichia coli cells, and the recombinant protein was purified to homogeneity. The optimal temperature and pH for Trehalose synthase activity were found to be $80^{\circ}C$ and 7.5, respectively. The half-life of heat inactivation was about 40 min at $90^{\circ}C$. The maximum trehalose conversion rate of maltose into trehalose by the enzyme increased as the substrate concentration increased, and reached 55.7% at the maltose concentration of 500 mM, implying that the enzyme conversion was dependent of the substrate concentration.

Chinese yam necrotic mosaic virus Isolated from Chinese Yam in Korea (한국산 장마(Dioscorea oppasita cv. Jang-Ma)에서 분리한 Chinese yam necrotic mosaic virus)

  • Kang, Dong-Kyoon;Kondo, Toru;Shin, Jong-Hee;Shin, Hye-Young;Sung, Jung-Hyun;Kang, Sang-Gu;Chang, Moo-Ung
    • Research in Plant Disease
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    • v.9 no.3
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    • pp.107-115
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    • 2003
  • Chinese yam (Dioscorea opposita cv. Jang-Ma) plants showing necrotic mosaic symptom were collected from their growing fields in Andong, Euisong, Gunwi and Daegu, Korea. Direct negative stainning method by electron microscope showed filamentous particles of about 660 nm in length. Immunosorbent electron microscopy (ISEM) revealed filamentous particles of 660nm decorated with antiserum of Chinese yam necrotic mosaic virus (ChYNMV). The virues purified partially were used to isolate viral RNA as template for RT-PCR to amplify about 1.2 kbp of 3'-terminal region (coat protein, 3'-UTR) with ChYNMV specific and oligo-dT primers. Amino acids sequences of amplified CP genes revealed that the viruses shared 97.9% similarity with ChYNMV (AB044386) wh ich was known as the member of Macluravirus. So the viruses from Chinese yam (D. opposita cv. Jang-Ma) plants were identified as ChYNMV. Comparing the CP amion acid sequences of ChYNMV strains with other macluraviruses such as Cardamon mosaic virus (CdMV), Narcissus latent virus (NLV) and Maclura mosaic virus (MacMV) revealed that N-terminal was the most varialbe region and conserved regions were present within the genus Macluravirus.

Molecular Cloning and mRNA Expression of the Bovine Peroxisome Proliperator Receptor Gamma(PPARγ) (한우 PPARγ 유전자의 동정과 mRNA의 발현)

  • Jeoung, Y.H.;Lee, S.M.;Park, H.Y.;Yoon, D.H.;Choi, J.G.;Moon, S.J.;Kang, M.J.
    • Journal of Animal Science and Technology
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    • v.46 no.1
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    • pp.23-30
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    • 2004
  • The peroxisome proliferator-activated receptor $\gamma$(PPAR$\gamma$), a member of the steroid/thyroid nuclear hormone receptor suferfamily of ligand-activated transcription factor, is an important regulator of adipocyte gene expression and differentiation. In this studies, we report the identification, characterization, and expression of a Hanwoo PPAR$\gamma$ gene. The PPAR$\gamma$ cDNA sequence of the Hanwoo show strong conservation with the corresponding sequences reported in other species except of three amino acid sequences. The distribution of PPAR$\gamma$ mRNA in various tissues of Korean cattle aged 12 months were investigated using Northern Blot analysis. The highest expression was detected in adipose tissue, more lower expression was detected in colon, small intestine, kidney, lung, while expression was not detected in brain, heart. PPAR$\gamma$ expression was higher in adipose tissue of Korean cattle when aged 30 months than aged 12 months. These results indicated PPAR$\gamma$, regulator adipocyte gene expression and differentiation, related on adipose differentiation in Korean native cattle(HANWOO).

(β-lapachone Regulates Tight Junction Proteins, Claudin-3 and -4, in Human Hepatocarcinoma Cells. (인체 간암세포에서 β-lapachone 처리에 의한 Tight Junction 관련 유전자의 변화)

  • Kim, Sung-Ok;Kwon, Jae-Im;Kim, Gi-Young;Kim, Nam-Deuk;Choi, Yung-Hyun
    • Journal of Life Science
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    • v.17 no.9 s.89
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    • pp.1298-1302
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    • 2007
  • A hallmark of cancers is 'leaky' tight junctions (Tjs). TJs mediated paracellular permeability is elevated and TJs maintained cell polarity is frequently lost. Concomitantly, TJs-associated proteins including members of the claudin family of proteins are dysregulated. Recent findings indicate that these TJs changes can contribute to cancer progression. In this study, we examined the effects of ${\beta}-lapachone$, a quinone compound obtained from the bark of the lapacho tree (Tabebuia avellanedae), on the Tjs-associated regulators in human hepatocarcinoma cell lines, HepG2 and Hep3B. ${\beta}-lapachone$ treatment downregulated the levels of insulin-like growth factor 1 receptor (IGF-lR) proteins in both HepG2 and Hep3B cells. But the levels of claudin-3 and -4 proteins were increased in ${\beta}-lapachone$-treated HepG2 and Hep3B cells. And also the zonnula occludens-l (la-I) and p-catenin protein levels by ${\beta}-lapachone$ were increased in a time-dependent manner. However, claudin-3 and -4 mRNA levels were uninhibited by ${\beta}-lapachone$ in HepG2 and Hep3B. The present results suggest that the upregulation of claudin-3 and -4 protein levels by ${\beta}-lapachone$ occurs by a post-transcriptional mechanism and points to a novel mechanism by ${\beta}-lapachone$.

DISTRIBUTION OF MUTANS STREPTOCOCCI IN DENTAL PLAQUE OF CHILDREN (소아의 치면세균막에 존재하는 mutans streptococci의 분포)

  • Kook, Joong-Ki;Park, Jong-Whi;Yoo, So-Young;Kim, Hwa-Sook;Lee, Nan-Young
    • Journal of the korean academy of Pediatric Dentistry
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    • v.31 no.3
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    • pp.439-447
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    • 2004
  • The aim of this study is to survey the frequency of mutans streptococci species and biotypes isolated from dental plaque in Korean children and the relationship between species and biotypes of mutans streptococci and dft index. Dental plaques were collected from the anterior and molar teeth of upper and lower jaws in the subjects, aged below 12 years old. A dental examination was performed for dft (decayed, filled, total) with the WHO caries diagnostic criteria. The mutans streptococci from the sample were cultured selectively on mitis salivarius-bacitracine (MSB) agar plate. For biotyping of mutans streptococci, biochemical test was performed. From the culture, bacterial genomic DNA was prepared for using of PCR template for the identification of mutans streptococci at the species-level. Forty strains of mutans streptococci were isolated from dental plagues of 40 patients. The biotype I (45%) and biotype IV (32.5%) were most frequently detected. The prevalence of S. mutans and S. sobrinus was 69% and 31%, respectively. There was no positive relationship between species and biotypes of mutans streptococci and dft index. Our results revealed that biotype I and S. mutans were frequently detected in Korean children and support that dental caries incidents by many causative factors not only bacterial factor.

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Identification and Characterization of Genes Involved in Cysteine Auxotrophy in Salmonella typhi (Salmonella typhi의 시스테인 영양요구성에 관여하는 유전자의 동정 및 특성 연구)

  • Lee, Sang-Ho;Kim, Sam-Woong;Yu, Jong-Earn;Yoo, Ah-Young;Kim, Young-Hee;Oh, Jeong-Il;Baek, Chang-Ho;Kang, Ho-Young
    • Journal of Life Science
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    • v.18 no.11
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    • pp.1507-1512
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    • 2008
  • In spite of long research period for Salmonella typhi, little information is known about the pathogenesis mechanism of human typhoid fever caused by S. typhi due to lack of infection model in animals. A wild-type of S. typhi Ty2 strain requires cysteine to grow on minimal media. We hypothesized that this cysteine requirement may restrict colonization of S. typhi in animals during infection process. Among the S. typhi strains carrying Salmonella typhimurium genomic library, we have isolated three S. typhi transformants growing on minimal media without cysteine. Although there were three ORFs in DNA of pBP71, the STM1490 ORF complemented cysteine auxotrophy of S. typhi. Analysis of the deduced amino acid sequence of the STM1490 homolog in S. typhi revealed that there are differences in two amino acids. Plasmids containing amino acid substitutions in STM1490 supported S. typhi growth on minimal media without cysteine, indicating irrelevance of these two amino acids to STM1490 function. These results tells us that there are other factors or systems involved in cysteine requirement of S. typhi.