• Title/Summary/Keyword: IS-PCR

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Detection of Novel Genetic Variations of the MG1R * 3 Allele in Pig(Sus scrofa) (돼지 Melanocortin Receptor 1(MC1R) 대립유전자 3의 신규 유전변이 탐색)

  • Cho, I.C.;Jeong, Y.H.;Jung, J.K.;Seong, P.N.;Oh, W.Y.;Ko, M.S.;Kim, B.W.;Lee, J.G.;Jeon, J.T.
    • Journal of Animal Science and Technology
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    • v.46 no.1
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    • pp.1-6
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    • 2004
  • This study was conducted to investigate novel genetic variations of MCIR^*3 allele. In general, white spotting or white belt on a black backgroud in pigs is determined by the E$^p$ allele at the MCIR/Extention locus. E$^p$ shares a frameshift mutation with the E$^{D2}$ allele for dominant black color. An oligonucleotide primer set was designed to amplify complete coding sequence of the porcine MCIR gene. The MCIR coding sequences obtained from five breeds those were Landrace(white). Yorkshire(white), Hampshire(belt), Berkshire(spot) and Jeju native black pigs(black), were used for this study. A multiple sequence alignment of the MCIR coding region using Clustal W was performed. The total length of the MCIR coding sequence ranged from 963 to 966 base pairs(bp) among the selected breeds. The sequence analysis of the complete coding region of MCIR was revealed that Hampshire and Jeju native black pig have 3 cytosines deletion and Birkshire has 2 cytosines deletion at codon 23(nt68) in Extention loci. Besides the finding, there were three different missense mutations and a frameshift mutation in the MCIR coding region.

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).

Expression Pattern of Antioxidant Enzymes Genes in the Ventral Prostates of Rats Exposed to Procymidone and/or Testosterone after Castration

  • Lee, Jong-Geol;Yon, Jung-Min;Jung, Ki-Youn;Lin, Chunmei;Jung, A-Young;Lee, Beom-Jun;Yun, Young-Won;Nam, Sang-Yoon
    • Journal of Embryo Transfer
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    • v.26 no.4
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    • pp.265-270
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    • 2011
  • Procymidone is a fungicide with anti-androgenic properties widely used to protect fruits from fungal infection, which induces an excessive reactive oxygen species production in male reproductive organs. In this study, to clarify whether procymidone affect the cellular antioxidant system of prostate at onset of puberty, gene expression patterns of the representative antioxidant enzymes such as cytoplasmic glutathione peroxidase (GPx1), phospholipid hydroperoxide GPx (PHGPx), selenoprotein P (SePP), cytoplasmic copper/zinc superoxide dismutase (SOD1), and manganese SOD (SOD2) were investigated in the rat ventral prostates exposed to procymidone using real-time RT-PCR analyses. Seven-week-old Sprague-Dawley rats castrated at 6 weeks old were treated with procymidone (25, 50, or 100 mg/kg per day) orally for 7 consecutive days after testosterone propionate (0.4 mg/kg per day) administration by subcutaneous injection. As compared to normal control animals, GPx1 mRNA expression in prostates significantly increased by the administration with TP and/or procymidone. However, PHGPx and SOD1 mRNA levels significanatly decreased by over 25 mg/kg of procymidone treatment and SePP and SOD2 mRNA levels was significanatly reduced by over 50 mg/kg of procymidone treatment. These findings indicate that procymidone may affect the antioxidant system of prostatic cells in up-regulation mode of GPx1, but in down-regulation modes of PHGPx, SePP, SOD1, and SOD2, suggesting that procymidone may affect differently the cellular antioxidant system of prostate according to the exposure doses.

Expression of Membrane Fusion Related Genes in Mouse Ovary (마우스 난소에서 막융합 관련 유전자의 발현)

  • Jung, Bok-Hae;Sung, Hyun-Ho;Park, Chang-Eun
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.1
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    • pp.8-14
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    • 2016
  • Granulosa cells surround the oocyte within the ovarian follicle and play an essential role in creating conditions required for oocyte as well as follicular development. The current study was conducted to examine the gene expression profile of mouse ovaries during the primordial to primary follicle transition process. Total RNAs from mouse ovaries on day 5 and day 12 were synthesized cDNA using annealing control primers. The DEGs were cloned and their identities were analyzed by BLAST search. The Plekha5 and Anxa11 were highly expressed in primary follicle stage. By contrast, their expression was increased in granulosa cells at the primary follicle stage. We have successfully discovered a list of genes expressed in day 5 and day 12 ovaries and confirmed that some of them are differentially expressed in PMF and/or PRI. This is a spatial-temporal regulatory mechanism on the ovarian folliculogenesis through membrane fusion. The gene expression profile from the current study would provide insight for future study on the mechanism(s) involved in primordial-primary follicular transition. This will provide information for identification of the mechanism of ovarian dysfunction.

The Prevalence of Plasmid-Mediated Quinolone Resistance Genes among CTX-M-14 Producing Escherichia coli Strains Isolated from a University Hospital in the Chungcheong Province (충청지역에 위치한 일개의 대학병원에서 분리된 CTX-M-14형 ESBL 생성 대장균을 대상으로 PMQR 유전자 빈도조사)

  • Sung, Ji Youn
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.3
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    • pp.210-216
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    • 2016
  • Recently, there has been a considerable increase in the prevalence of CTX-M type extended-spectrum ${\beta}$-lactamase (ESBL)-producing E. coli isolates worldwide, including Korea. To investigate the ESBL genes in the E. coli strains isolated from a university hospital in the Chungcheong area, a study was conducted using PCR amplification and nucleotide sequence analysis of the amplified products to detect the plasmid mediated quinolone resistance (PMQR) genes in ESBL producing E. coli isolates. The number of CTX-M-14 producing isolates was 25 (16.0%) isolates, and of them, 9 (5.8%) isolates also produced CTX-M-15. All CTX-M type ESBL producing E. coli isolates showed resistance to cefotaxime. Twelve (48%) CTX-M type ESBL producing E. coli isolates contained the PMQR genes, 8 contained qnrS1, and 8 contained aac(6')-Ib-cr. Four isolates harbored both qnrS1 and aac(6')-Ib-cr genes. In our study, we confirmed that the plasmid mediated antimicrobial resistant determinants-the ESBL and PMQR genes-were distributed in the E. coli isolates. To prevent further spreading of the resistant genes among the E. coli isolates, consistent effort is required to investigate and monitor the resistant genes.

Antimicrobial Effect of Extract of Glycyrrhiza uralensis on Methicillin-Resistant Staphylococcus aureus (감초 추출물이 항생제 내성균주의 항균활성에 미치는 영향)

  • Lee, Ji-Won;Ji, Young-Ju;Yu, Mi-Hee;Im, Hyo-Gwon;HwangBo, Mi-Hyang;Lee, In-Seon
    • Korean Journal of Food Science and Technology
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    • v.37 no.3
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    • pp.456-464
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    • 2005
  • Antimicrobial drug-resistance is natural response to antimicrobial stress based on selection, which weakens chemotherapy effect. Introduction of large numbers of chemotherapeutic agents to clinical practice has generated strains of microorganisms that survive and multiply in vivo with high-drug concentrations. Methicillin-resistant Staphylococcus aureus (MRSA), bacteria found in normal daily life, can be easily ingested through milk vegetables, and meats, etc. MRSA emerged in many port of the world, increasing complex clinical problems. Therefore, new agents are needed to treat MRSA. Glycyrrhiza uralensis was extracted using 80% MeOH to investigate its antimicrobial activity against MRSA stains KCCM 11812, 40510, and 40512 through bacterial measurement, disc diffusion, and O.D. methods, MIC values, MRSA gene expression investigation, and scanning electron microscope observation. Results revealed MecA, Mecl, MecRI, and FemA were the most highly manifested MRSA genes. Methanolic extract of G. uralensis significantly inhibited MRSA and thus could be used in development of antibacteria.

Therapeutic effects of Hominis placenta herb-acupuncture in adjuvant-induced arthritis rat (흰쥐의 Adjuvant 관절염에 대한 자하거(紫河車) 약침의 효과)

  • Yeom, Mi-Jung;Kang, Ji-Eun;Hahm, Dae-Hyun;Park, Hi-Joon;H.Lee, Eun-Joo;Shim, In-Sop;Lee, Hye-Jung
    • Journal of Pharmacopuncture
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    • v.5 no.1 s.8
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    • pp.91-103
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    • 2002
  • Rheumatoid arthritis (RA) is a chronic inflammatory autoimmune disease. characterized by leukocyte infiltration, a chronic inflammation of the joint, a pannus formation and the extensive destruction of the 3Iticular caJ1ilage and bone. Several proinflammatory cytokines such as tumor necrosis factor-${\alpa}$(TNF-${\alpa}$), interleukin-1${\beta}$ (IL-1${\beta}$) and interleukin 6 (IL-6) have been implicated in the pathological mechanisms of synovial tissue proliferation, joint destruction and programmed cell death in rheumatoid joint. In the Korean traditional medicine, Hominis placenta (HP) as an herbal solution of herb-acupuncture has been widely used to treat the inflammatory diseases including RA. In order to study the medicinal effect of HP herb-acupuncture on rheumatoid joint, an adjuvant-induced arthritis (AlA) was generated by the injection of 1.5 mg uf Mycobactelium tuberculusis. emulsified in squalene, 10 the base of the tail of Sprague-Dawley(SD) rats. After onset stage of polyarthritis, HP was daily injected to the Zusanti (ST36) acupuncture points in both of rat lags and the expression pattems of cytokines such as TNF-{\alpa}$, IL-1${\beta}$, and 1L-6 at the knee joint were analyzed using immunostaining and RT-PCR. The HP herb-acupuncture was found to be effective to alleviate the arthritic symptums in adjuvant-induced arthritic rats as regards the joint appearance and the expression profiles of inflammatory cytokines. In conclusion, therapeutic effects of HP herb-acupuncture on the rat with AlA might be related to anti inflammatory activities of the hurb-acupuncture.

Molecular Analysis of Isoniazid-Resistance Related Genes of Mycobacterium tuberculosis Isolated from Korea

  • Hwang Joo Hwan;Jeong Eun Young;Choi Yeon Im;Bae Kiho;Song Taek Sun;Cho Sang-Nae;Lee Hyeyoung
    • Biomedical Science Letters
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    • v.11 no.4
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    • pp.455-463
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    • 2005
  • Resistance to isoniazid (INH), which is one of the most important drugs in Mycobacterium tuberculosis chemotherapy, has been associated with mutations in genes encoding the mycobacterial catalse-peroxidase (katG), the enoyl acyl carrier protein (ACP) reductase (inhA), alkyl hydroperoxide reductase (ahpC), beta-ketoacyl acyl carrier protein synthase (kasA), and NADH dehydrogenase (ndh). In this study, we examined INH-resistance related genes in 50 INH-resistant and 24 INH-susceptible isolates by PCR-sequence analysis. In brief, mutations at the katG gene were found at codon 315 alone (2/50), at codon 463 alone (19/50), and both at 315 and 463 (29/50). However, while mutations at codon 315 were only detected in INH-resistant isolates, mutations at codon 463 were also detected in INH-susceptible isolates indicating mutations at 463 alone do not seem to confer resistance to INH. Similar to the case of katG 463, some of inhA mutations were also found among INH-susceptible isolates. For example, whereas mutations at 8 upstream of the start codon (UPS) and 15 UPS of the inhA gene were detected only in INH-resistant isolates, mutations at 101, 115, and 125 UPS were detected only in INH-susceptible isolates. Many different kinds of mutations were detected in INH­resistant isolates at ahpC, oxyR gene, and intergenic region of the oxyR-ahpC genes. Howerver, the mutations were not found oxyR and the intergenic regions in INH-susceptible isolates. No mutations were found at either kasA or at ndh gene among INH-resistant isolates. In conclusion, some of mutations such as katG 315, inhA promotor region, and oxyR-ahpC seem to be strongly related to INH-resistance. Currently we are developing a molecular diagnostic method based on these results.

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Association between Tardive Dyskinesia and T103C Polymorphisms of 5-$HT_{2A}$ Receptor Gene (지연성 운동장애와 5-$HT_{2A}$ 수용체 유전자 T103C 다형성과의 관계)

  • Hahn, Sang Woo;Shin, Jeong Won;Choi, Tae Youn;Woo, Sung Il;Jung, Han Yong;Jung, Hee Yeoun;Han, Sun Ho
    • Korean Journal of Biological Psychiatry
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    • v.10 no.2
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    • pp.133-140
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    • 2003
  • Objective:Some candidate gene polymorphisms were reported to be associated with tardive dyskinesia (TD). The aim of this study was to investigate the association of the 5-$HT_{2A}$ receptor gene polymorphisms with TD in Korean schizophrenic subjects. Method:Subjects were of 59 schizophrenic patients with TD and 60 schizophrenic patients without TD for studying of 5-$HT_{2A}$ receptor gene polymorphisms. TD was evaluated using the Abnormal Involuntary Movement Scale(AIMS). Genomic DNA was amplified by PCR and digestion with MspI and BsmI. Result:There were no statistically significant differences in the demographic variables, such as age, male to female percentage, duration of illnesses and duration of antipsychotic drug exposure between the TD group and control group. 1) T102C polymorphisms and TD Comparing the TD group and control group, the 102T/C allele was associated with a significantly increased risk for TD (${\chi}^2$=5.560, df=1, p=0.018). 2) Three AIMS categories of TD and T102C genotype. There were statistically significant differences in the three AIMS categories(${\chi}^2$=6.835, df=2, p=0.033). Conclusion:These result suggest 102T/C genotypes of the 5-$HT_{2A}$ receptor gene are related to the development of TD. The 102T/C genotypes were associated with significantly higher AIMS orofacial dyskinesia scores. These findings suggest that the 5-$HT_{2A}$ receptor gene is significantly associated with susceptibility to TD in patients with chronic schizophrenia.

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Evaluation of the Accuracy of the $EasyTest^{TM}$ Malaria Pf/Pan Ag, a Rapid Diagnostic Test, in Uganda

  • Chong, Chom-Kyu;Cho, Pyo Yun;Na, Byoung-Kuk;Ahn, Seong Kyu;Kim, Jin Su;Lee, Jin-Soo;Lee, Sung-Keun;Han, Eun-Taek;Kim, Hak-Yong;Park, Yun-Kyu;Cha, Seok Ho;Kim, Tong-Soo
    • Parasites, Hosts and Diseases
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    • v.52 no.5
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    • pp.501-505
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    • 2014
  • In recent years, rapid diagnostic tests (RDTs) have been widely used for malaria detection, primarily because of their simple operation, fast results, and straightforward interpretation. The Asan $EasyTest^{TM}$ Malaria Pf/Pan Ag is one of the most commonly used malaria RDTs in several countries, including Korea and India. In this study, we tested the diagnostic performance of this RDT in Uganda to evaluate its usefulness for field diagnosis of malaria in this country. Microscopic and PCR analyses, and the Asan $EasyTest^{TM}$ Malaria Pf/Pan Ag rapid diagnostic test, were performed on blood samples from 185 individuals with suspected malaria in several villages in Uganda. Compared to the microscopic analysis, the sensitivity of the RDT to detect malaria infection was 95.8% and 83.3% for Plasmodium falciparum and non-P. falciparum, respectively. Although the diagnostic sensitivity of the RDT decreased when parasitemia was ${\leq}500\;parasites/{\mu}l$, it showed 96.8% sensitivity (98.4% for P. falciparum and 93.8% for non-P. falciparum) in blood samples with parasitemia ${\geq}100\;parasites/{\mu}l$. The specificity of the RDT was 97.3% for P. falciparum and 97.3% for non-P. falciparum. These results collectively suggest that the accuracy of the Asan $EasyTest^{TM}$ Malaria Pf/Pan Ag makes it an effective point-of-care diagnostic tool for malaria in Uganda.