• Title/Summary/Keyword: RFLP.

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Two new genotypes of Plasmodium vivax circumsporozoite protein found in the Republic of Korea

  • Kho, Weon-Gyu;Park, Yeong-Hong;Chung, Joon-Yong;Kim, Jong-Pil;Hong, Sung-Tae;Lee, Won-Ja;Kim, Tong-Soo;Lee, Jong-Soo
    • Parasites, Hosts and Diseases
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    • v.37 no.4
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    • pp.265-270
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    • 1999
  • The gene encoding Plasmodium vivax circumsporozoite protein (PvCSP) exhibits polymorphism in many geographical isolates. The present study was designed to investigate polymorphism in PvCSP gene of P. vivax isolates in Korea. Thirty isolates, obtained from indigenous cases in Yonchon-gun, Kyonggi-do in 1997, were subjected for sequencing and RFLP analysis of the repeat and post-repeat regions of PvCSP gene and two genotypes (SK-A and SK-B) were identified. The genotype of 19 isolates was SK-A and that of 11 isolates was SK-B. Although the number of 12-base repeats present in SK-A was three while two were found in a Chinese strain CH-5, the repeat sequence of SK-A was identical to that of CH-5 except for one base substitution. Compared with known data there was no identical isolates with SK-B, but the sequence of SK-B was similar to that of a North Korean (NK) isolate. These results indicate that two genotypes of PvCSP coexist in the present epidemic area of Korea and the present parasite may originate from East Asia. RFLP would be useful to classify genotypes of P. vivax population instead of gene sequencing.

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Four Cases of Taenia saginata Infection with an Analysis of COX1 Gene

  • Cho, Jaeeun;Jung, Bong-Kwang;Lim, Hyemi;Kim, Min-Jae;Yooyen, Thanapon;Lee, Dongmin;Eom, Keeseon S.;Shin, Eun-Hee;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • v.52 no.1
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    • pp.79-83
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    • 2014
  • Human taeniases had been not uncommon in the Republic of Korea (=Korea) until the 1980s. The prevalence decreased and a national survey in 2004 revealed no Taenia egg positive cases. However, a subsequent national survey in 2012 showed 0.04% (10 cases) prevalence of Taenia spp. eggs suggesting its resurgence in Korea. We recently encountered 4 cases of Taenia saginata infection who had symptoms of taeniasis that included discharge of proglottids. We obtained several proglottids from each case. Because the morphological features of T. saginata are almost indistinguishable from those of Taenia asiatica, molecular analyses using the PCR-RFLP and DNA sequencing of the cytochrome c oxidase subunit 1 (cox1) were performed to identify the species. The PCR-RFLP patterns of all of the 4 specimens were consistent with T. saginata, and the cox1 gene sequence showed 99.8-100% identity with that of T. saginata reported previously from Korea, Japan, China, and Cambodia. All of the 4 patients had the history of travel abroad but its relation with contracting taeniasis was unclear. Our findings may suggest resurgence of T. saginata infection among people in Korea.

Genotype analysis of Cryptosporidium spp. prevalent in a rural village in Hwasun-gun, Republic of Korea

  • PARK Jae-Hwan;GUK Sang-Mee;HAN Eun-Taek;SHIN Eun-Hee;KIM Jae-Lip;CHAI Jong-Yil
    • Parasites, Hosts and Diseases
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    • v.44 no.1 s.137
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    • pp.27-33
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    • 2006
  • Two species of Cryptosporidium are known to infect man; C. hominis which shows anthroponotic transmission between humans, and C. parvum which shows zoonotic transmission between animals or between animals and man. In this study, we focused on identifying genotypes of Cryptosporidium prevalent among inhabitants and domestic animals (cattle and goats), to elucidate transmittal routes in a known endemic area in Hwasun-gun, Jeollanam-do, Republic of Korea. The existence of Cryptosporidium oocysts was confirmed using a modified ZiehlNeelsen stain. Human infections were found in 7 $(25.9\%)$ of 27 people examined. Cattle cryptosporidiosis cases constituted 7 $(41.2\%)$ of 17 examined, and goat cases 3 $(42.9\%)$ of 7 examined. Species characterizations were performed on the small subunit of the rRNA gene using both PCR-RFLP and sequence analysis. Most of the human isolates were mixtures of C. hominis and C. parvum genotypes and similar PCR-RFLP patterns were observed in cattle and goat isolates. However, sequence analyses identified only C. hominis in all isolates examined. The natural infection of cattle and goats with C. hominis is a new and unique finding in the present study. It is suggested that human cryptosporidiosis in the studied area is caused by mixtures of C. hominis and C. parvum oocysts originating from both inhabitants and domestic animals.

Identification of MC1R gene variants of Hanwoo and Holstein meat using PCR-RFLP (PCR-RFLP를 이용한 한우와 젖소고기의 MC1R 유전자변이 검출)

  • Koh Ba-Ra-Da;Kim Yong-Hwan;Park Seong-Do;Na Ho-Myung;Kim Jeong-Nam;Sung Chang-Min;Lee Sam-Soo
    • Korean Journal of Veterinary Service
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    • v.28 no.3
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    • pp.259-265
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    • 2005
  • The melanocortin 1 receptor (MC1R) encoded by the coat color extension gene (E) plays a key role in the signaling pathway of melanin synthesis. The primers for the amplification of bovine MC1R gene were designed based on a bovine MC1R gene sequence (GenBank accession no. Y19103). A size of 483bp (482bp for Hanwoo) was amplified by PCR, digested with Hpa II restriction enzyme and electrophoresed in $1.5\%$ agarose gel. When the amplified DNA product (483 bp) was digested with Hpa II restriction enzyme, Hanwoo meat showed a single band of 482bp, whereas two fragments of 325bp and 158bp were detected in Holstein, Angus and meat of Hanwoo / Holstein cross cow having back coat color phenotype, respectively. The results of this experiment Indicate that new designed primers of bovine MCIR gene may be useful for identification of Hanwoo meat from Holstein, Black Angus and Hanwoo / Holstein cross cow meat.

BRAFV600E Mutation Analysis in Fine Needle Aspiration Biopsy Cytology and Formalin Fixed Paraffin Embedding Block of the Thyroid

  • Han, Kyung Hee;Park, Won Young;Lee, Young Nam
    • Korean Journal of Clinical Laboratory Science
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    • v.45 no.2
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    • pp.66-72
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    • 2013
  • Fine Needle Aspiration Biopsy Cytology (FNABC), which is known as the most accurate and cost-effective method for diagnosis of the thyroid nodule, may still result in indeterminate cases that are cellular paucity and show minor nuclear atypia. However, most cases are associated with suspicion of papillary thyroid carcinoma (PTC). A B-type Raf kinase (BRAF) mutation was found in about half of PTCs which is currently helping us to differentiate malignancies from benign lesions. Cases studied included 46 histological, confirmed PTC cases. FNABC 102 cell paucity and 74 atypia benign cases were previously diagnosed as suspicious of PTC using cytologic examination. These cases were analyzed for BRAF mutation by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) with a new restriction enzyme. In this study, the sensitivity and specificity were calculated and, BRAF mutation was detected by means of a histological method in 23 of 46 cases of PTC and no mutation was found in 22 cases. However, one case was not detected. In using FNABC, BRAF mutation was detected in 6 of 102 cases in cell paucity and in 11 of 74 cases in the atypia. Two cases were not detected in the atypia. The sensitivity and specificity of PCR-RFLP in FNABC were 60% and 97.4% respectively. Assessment of Formalin Fixed Paraffin Embedding (FFPE) block demonstrated similarly a 51.1% positive and 48.9% negative in PTC. Evaluation of BRAF mutation revealed high specificity and low sensitivity in using FNABC method. This study suggests that BRAF mutation analysis should be useful for the clinical diagnosis of PTC in FNABC with cytological findings suspicious for PTC.

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Microbial Community in the TPH-Contaminated Aquifer for Hot Air Sparging using Terminal-Restriction Fragment Length Polymorphism (유류오염대수층 고온공기분사공정시 제한효소다형성 미생물 군집)

  • Lee, Junho;Park, Kapsong
    • Journal of Korean Society on Water Environment
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    • v.24 no.1
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    • pp.19-29
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    • 2008
  • Hot air sparging is a groundwater remediation technique, in which organic contaminants volatilized into hot air from the saturated to vadose zone. In the laboratory diesel (10,000 mg TPH/kg) was spiked in contaminated saturated aquifer soil. The hot air ($34.9{\pm}2.7^{\circ}C$) was injected in intermittent (Q=1,500 mL/min, 10 minute injection and 10 minute idle) modes. We performed microcosm tests using the groundwater samples to assess TPH reductive remediation activity. For Terminal-Restriction Fragment Length Polymorphism (T-RFLP) analysis of eubacterial communities in sludge of wastewater treatment plants and soil of experiment site, the 16S rDNA was amplified by Polymerase Chain Reaction (PCR) from the sludge and the soil. The obtained 16S rDNA fragments were digested with Msp I and separated by electrophoresis gel. We found various sequence types for hot air sparging experiment with sludge soil samples that were closely related to Bacillus (149 bp, Firmicutes), Methlobacterium (149 bp, Euryarchaeotes), Pseudomonas (492 bp, ${\gamma}$-Proteobacteria), etc., in the clone library. In this study we find that TPH-water was reduced to 78.9% of the initial value in this experiment aquifer. The results of the present study suggests that T-RFLP method may be applied as a useful tool for the monitoring in the TPH contaminated soil fate of microorganisms in natural microbial community.

SNP Analysis of the Human LDL Receptor and CETP Gene in Korean Subjects with Hypertension

  • Shin, Jae-Hyun;Kang, Byung-Yong;Lee, Kang-Oh;Eo, Hyun-Shun;Bae, Joon-Seol;Kim, Ki-Tae;Kim, Hyun-Sup;Lee, Chung-Choo
    • Environmental Mutagens and Carcinogens
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    • v.22 no.1
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    • pp.47-53
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    • 2002
  • Essential hypertension is complex disorder influenced by multiple genetic and environmental factors. Alterations of lipid metabolism in plasma have been reported to be related to an increased risk of essential hypertension. The aim of this study was to investigate the relationship between two SNPs of the human LDL receptor and CETP gene and hypertension in Korean population. There were no significant differences in allele and genotype frequencies of two SNPs in normotensives and hypertensives. With respect to Hinc II RFLP in the LDL receptor gene, pooled odds ratio value indicated the significant heterogeneity among populations studied by meta-analysis (Breslow-Day test df = 2, P<0.05). In the case of Bam HI RFLP in the CETP gene,. our study is the first report of an association between the SNP of the CETP gene and hypertension, although our result failed to demonstrate the significant association between the Bam HI RFLP of the CETP gene and hypertension in Korean population. Further work, using larger sample sizes and various ethnic groups, is required to establish the precise role of these two candidate gene polymorphisms on hypertension.

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Variation in trn-L/trn-V and trn-F/trn-T spacer regions of cpDNA in Abies koreana Wilson and A. nephrolepis Traut./Maxim

  • Kormutak, A.;Hong, Y.-P.;Kwon, H.-Y.;Kim, C.-S.
    • Journal of Korean Society of Forest Science
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    • v.96 no.2
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    • pp.131-137
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    • 2007
  • The first evidence has been provided about the variation within trnL-trnV and trnF-trnT spacer regions of cpDNAs in Korean fir and Manchurian fir, revealed by PCR-RFLP analysis. Four cpDNA haplotypes have accordingly been recognized by being analyzed using the trnL-trnV/Tru11 primer-enzyme combination and 3 haplotypes using the trnF-trnT/TagI combination, which exhibited inter and intraspecific variation. A total of 6 cpDNA haplotypes were recognized by pooling the PCR-RFLP variants observed in both combinations. Haplotypes 2 and 3 were common for both species investigated, whereas haplotypes 1, 4, and 5 were detected only in Korean fir and haplotype 6 was detected only in Manchurian fir. Although haplotypes 2 and 3 were common in both species, haplotype 2 was major haplotype for Korean fir and haplotype 3 was one of the 2 major haplotypes for Manchurian fir. Restricted occurrence of haplotype 4 in Mt. Halla and haplotype 5 in Mt. Jiri of the Korean fir may represent the existence of geographic isolation by the sea between them. Diagnostic potential of individual haplotypes in discriminating between the two species as well as between their populations is discussed.

Identification of Korean Native Pork Using Breed-Specific DNA Marker of KIT Gene

  • Chung, Eui-Ryong;Chung, Ku-Young
    • Food Science of Animal Resources
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    • v.30 no.3
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    • pp.403-409
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    • 2010
  • Accurate methods for the identification of closely related species or breeds in raw and processed meats must be developed in order to protect both consumers and producers from mislabeling and fraud. This paper describes the development of DNA markers for the discrimination and improvement of Korean native pig (KNP) meat. The KIT gene is related to pig coat color and is often used as a candidate marker. A 538 bp fragment comprising intron 19 of the pig KIT gene was amplified by PCR using specific primers, after which the PCR amplicons of a number of meat samples from KNP and three major improved breeds (Landrace, Duroc and Yorkshire) were sequenced in order to find a nucleotide region suitable for PCR-RFLP analysis. Sequence data showed the presence of two nucleotide substitutions, g.276G>A and g.295A>C, between KNP and the improved pig breeds. Digestion of KIT amplicons with AccII enzyme generated characteristic PCR-RFLP profiles that allowed discrimination between meats from KNP and improved pig. KNP showed three visible DNA bands of 264/249, 199, and 75 bp, whereas DNA bands of 249, 199, and 90 bp were detected in the three improved pig breeds. Therefore, the 75 bp DNA fragment was specific only to KNP, whereas the 90 bp DNA fragment was specific to the improved breeds. The breed-specific DNA markers reported here that target the KIT gene could be useful for the identification of KNP meat from improved pig meats, thus contributing to the prevention of falsified breed labeling.

Studies on the MC1R Gene Frequencies in Landrace, Large White, Duroc and Jeju Native Black Pigs (랜드레이스, 대요크셔, 듀록 및 제주 흑돈의 Melanocortin 1 Receptor(MC1R) 유전자의 유전자형 분석)

  • Cho, I.C.;Lee, J.G.;Jung, J.G.;Yang, B.S.;Kang, S.Y.;Kim, B.W.
    • Journal of Animal Science and Technology
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    • v.44 no.2
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    • pp.207-212
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    • 2002
  • This study was conducted to investigate the genotypes and frequencies of Melanocortin 1 Receptor(MC1R) genes in pigs which plays a central role in regulation of eumelanin (black/brown) and phaeomelanin(red/yellow) pigment synthesis within the mammalian melanocytes. Four different breeds of pigs(20 Landrace, 20 Yorkshire, 20 Duroc, and 93 Jeju native black pigs) were used and PCR-RFLP analysis of MC1R gene was also carried out. Two regions of MC1R genes (428bp and 405bp) were amplified using two specific primers (MERL1-EPIG2, EPIG1-EPIG3), respectively and MC1R allele were determined using 2 restriction enzymes (BspHⅠ, AccⅡ). The results of this experiment indicated that MC1R allelic type in Landrace, Large Yorkshire and Duroc were MC1R *2 (Ep), MC1R *2 (Ep), MC1R *4 (e), respectively. However, various allelic types of MC1R genes were detected in Jeju native black pigs. MC1R allelic type of Jeju black pigs was MC1R*2 type as in Meishan and Large black breeds or MC1R*3 type as in Hampshire and Berkshire breeds and the gene frequencies of ED1 and ED2 were 0.554 and 0.446 in average.