• Title/Summary/Keyword: MspA1I

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Mixed Infection of 16S rDNA I and V Groups of Phytoplasma in a Single Jujube Tree

  • Lee, Sang-Hun;Han, Sang-Sub;Cha, Byeong-Jin
    • The Plant Pathology Journal
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    • v.25 no.1
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    • pp.21-25
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    • 2009
  • Jujube trees infected with phytoplasma exhibit symptoms of typical witches' broom, such as yellowing, abnormally small leaves, short internodes and proliferation of shoots. A 1.2 kb fragment of the 16S rDNA from jujube phytoplasma was generated by R16F2n/R16R2 primer pair from earlier amplified P1/P7 PCR products of cloned jujube witches' broom phytoplasmas. Enzymatic restriction fragment length polymorphism (RFLP) and sequence analysis of 16S rDNA revealed that the jujube tree was infected with 16S rDNA I and V groups of phytoplasmas. Extensive comparative analyses of restriction enzyme profiles from Alu I, Hha I, Msp I, and Rsa I clearly classified the two into different phytoplasma groups. The phylogenie analyses based on 16S rDNA showed that the similarity of the two different clones was 87.5%. This is the first report of a mixed phytoplasmal infection in a single jujube tree.

Cloning and Base Sequence Determination of Replication Initiation Gene (rep) Isolated from Staphylococcus aureus DH1 R-plasmid pSBK203 (Staphylococcus aureus DH1에서 분리된 R-plasmid pSBK203의 복제 개시 유전자(rep) 분리 및 염기서열 결정)

  • Park, Seung-Moon;Kwon, Dong-Hyun;Byeon, Woo-Hyeon
    • Korean Journal of Microbiology
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    • v.31 no.1
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    • pp.44-47
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    • 1993
  • A replication initiation gene was identified and its nucleotide sequence has been determined from a 3.8 kb, chloramphenicol acethyltransferase conferring R-plasmid pSBK203 of Staphylococcus aures. Location of the replication related region of pSBK 203 was determined by interuption with pUC 119 at XBaI and MspI sites which resulted in inactivation of replication in Bacilius subtilis. Base sequence of this region revealed on open reading frame of 942 base pairs, which encoded a 314 amino acid protein. Base sequence homology with other rep of pT181 family plasmids such as pT181, pC221, pC223, pS194, pU112, and pCW7 was ranged from 78% to 97% and the predicted amino acid sequence homology was from 72% to 95%.

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Characterization of Cucumver mosaic virus Isolated from Hydrangea macrophylla for. otaksa (Sieb. et Zucc) Wils. (수국에서 분리한 Cucumber mosaic virus의 특성)

  • 방주희;박선정;이금희;최장경;이상용
    • Research in Plant Disease
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    • v.7 no.1
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    • pp.1-7
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    • 2001
  • An isolate of Cucumber mosaic cucumovirus(CMV) was isolated from Hydrangea macrophylla for. otaksa(Sieb. et Zucc. ) Wils. showing mosaic symptoms, and designated as Hm-CMV. Hm-CMV was characterized by the tests of host range, physical properties, serological properties, RNA and coat protein compositions, and reverse transcription and polymerase chain reaction (RT-PCR) analysis. Twelve species in 4 families were used in the host range test of Hm-CMV and could be differentiated from Y-CMV used as a control CMV by the ringspot and line pattern on inoculated leaves of several tobacco plants. Thevirus produced local lesions on inoculated leaves of Chenopodium amarticolor, C. quinoa and Vigna unguiculata. The physical properties of the virus were as follows; thermal inactivation point(TIP) was 60$\^{C}$, dilution end point (DEP) was 10$\^$-3/, and longevity in vitro (LIP) was 3∼4 days. Hm-CMV was serologically identical to Y-CMV. SDS-polyaciylamide gel electrophoresis(SDS-PAGE) showed one major protein band of about 28 kDa. In RNA or dsRNA analysis, Hm-CMV consisted of four RNA or dsRNA species, but satellite RNA was not detected. In RT-PCR using CMV-common primer and CMV subgroup I-specific primer, bothe amplified expected size of about 490 bp and 200 bp DNA fragments from Hm-CMV, respectively. Restriction enzyme analysis of the 490 bp RT-PCR products using EcoR I and Msp I showed that Hm-CMV belonged to CMV subgroup I. However, Hm-CMV could be differentiated from other CMV subgroup I isolates by RNA fingerprinting by arbitrarily primed polymerase chain reaction (RAP-PCR).

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PCR-mediated Fingerprinting to Identify Dang-Gui(당귀) (당귀류 한약재의 유전자 감별 연구)

  • 최호영;정유헌;고지완
    • The Journal of Korean Medicine
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    • v.20 no.4
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    • pp.11-15
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    • 2000
  • Radix Angelicae Gigantis is sweet and pungent in flavor, warm in property. Its effects are tonifying the blood, promoting blood circulation, relieving pain and moistening the bowels. Its indications are blood deficiency syndrome characterized by sallow complexion, dizziness, irregular menstruation, amenorrhea, pains due to blood stasis, and rheumatic arthralgia. Using genes of A. gigas, A. acutiloba, and A. sinensis, the origin of which is identified, as criteria, we analysed many kinds of Angelica with RAPD and RFLP on ITS region, in order to compare and discriminate genes extracted from crude drugs ‘Dang-gui’, that are produced in Korea on the one hand and imported on the other hand. We reached the following conclusion. 1. We could extract DNA from both original plant and dried plant. 2. Especially Uniprimer #1, Uniprimer #2, Uniprimer #4 and Uniprimer #9 were useful. 3. Among the restriction enzymes Sma I, Msp I, Hae III, and Hinf I, used in this experiment, four restriction enzymes except Hinf I could be used properly in discriminating all samples used as A. gigas. We think that this result can be used as a method of discriminating crude drug of Angelica L. related drugs, and used in controlling quality and circulation.

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Analysis of p53 and Retinoblasoma(Rb) Gene Polymorphisms in Relation to Lung Cancer in Koreans (한국인 폐암 환자에 대한 p53 및 Rb유전자의 다형성 분석)

  • Lee, Kyung-Sang;Sohn, Jang-Won;Yang, Suck-Chul;Yoon, Ho-Joo;Shin, Dong-Ho;Park, Sung-Soo;Lee, Jung-Hee;Lee, Chun-Geun;Cho, Youl-Hee
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.3
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    • pp.534-546
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    • 1997
  • Background : The p53 and retinoblastoma(Rb) tumor suppressor genes are associated with the pathogenesis of several types of human cancer. Substantial proportion of the primary lung cancers or cell lines have been reported to have the p53 and/or the Rb gene mutations. But, so far there is no report on the analysis of the Rb gene polymorphism as one of the genetic susceptibility marker. This study was undertaken to establish the gene frequencies of the polymorphic genotypes of the p53 and Rb genes in Koreans to evaluate the possible involvement of these genotypes as a risk factor of lung cancer. Methods : In this study 145 controls without previous and present tumor history and 128 lung cancer patients were subjected to analysis. The two intragenic polymorphisms of the p53 gene(exon 4/ AccII, intron 6/MspI) and one intron 17/XbaI polymorphism of the Rb gene were analysed by the method of polymersae chain reaction- restriction fragment length polymorphisms(PCR-RFLPs). The genotype of the intron 3/16 bp repeat polymorphism of p53 was determined by PCR and direct gel electrophoresis. Results : There were no significant differences in the genotype distributions of the p53 gene between lung cancer patients and controls. But heterozygotes(Arg/Pro) of the exon 4/AccII polymorphisms were slightly over-represented than controls, especially in the Kreyberg type I cancer, which was known to be associated with smoking. The intron 3/16 bp duplication and the intron 6/MspI polymorphisms were in complete linkage disequilibrium. About 95% of the individuals were homozygotes of the common alleles both in the 16 duplication and MspI polymorphisms, and no differences were deteced in the genotype distributions between lung cancer patients and controls. Overall genotype distributions of the Rb gene polymorphisms between lung cancer patients and controls were not significantly different However, the genotype distributions in the Kreyberg type I cancer were significantly different from those of controls(p = 0.0297) or adenocarcinomas(p = 0.0008). It was noticeable that 73.4% of the patients with adenocarcinomas were heterozygotes(r1/r2) whereas 39.2% of the Kreyberg type I cancer were heterozygous at this polymorphisms. In the lung cancer patients, significant differences were also noted between the high dose smokers and low dose smokers including non-smokers(p = 0.0258). The relative risk to Kreyberg type I cancer was significantly reduced in the individuals with the genotype of r1/r2(odds ratio = 0.46, 95% C.I. = 0.25-0.86, p = 0.0124). The combined genotype distribution of the exon 4 AccII of the p53 and the intron 17 Rb gene polymorphisms in Kreyberg type I cancers were significantly different from dose of controls or adenocarcinomas. The highest odds ratio were observed in the individuals with the genotypes of Arg/Pro and r2/r2(odds ratio = 1.97,95% C.I. = 0.84-4.59) and lowest one was in the patients with Arg/Arg, r1/r2 genotype(odds ratio = 0.54, 95% C.I. = 0.25-1.14). Conclusion : The p53 and the Rb gene polymorphisms modulate the risk of smoking induced lung cancer development in Koeans. However, the exact mechanism of risk modulation by these polymorphism remains to be determined. For more discrete clarification of associations between specific genotypes and lung cancer risk, the evaluations of these polymorphisms in other ethnics and more number of patients will be needed.

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Electrochemical Characteristics of EDLC with various Organic Electrolytes (유기전해질에 따른 EDLC의 전기화학적 특성)

  • Yang Chun-Mo;Lee J.K.;Cho W.I.;Cho B.W.;Rim Byung-O
    • Journal of the Korean Electrochemical Society
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    • v.4 no.3
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    • pp.113-117
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    • 2001
  • Specific capacitance and charge-discharge rate of EDLC using activated carbon electrode were affected by the compositions of electrolytes, the conditions of charge-discharge and physical properties of activated carbon materials. The activated carbon electrode was prepared by dip coating method. Charge-discharge test and electrochemical experiments were carried out for various kinds of organic electrolytes. Effects of charge and discharge current density on the specific capacitance were studied. Characteristics of leakage current, self-discharge and time-voltage curves in optimum conditions of organic electrolytes were compared with conventional $1M-Et_4NBF_4/PC$ electrolyte. The EDLC using MSP-20(specific surface area: $2000m^2/g$) electrode and $1M-LiPF_6/PC-DEC(1:1)$ was exhibited th highest specific capacitance of 130F/g and low polarization resistances. The EDLC using MSP-20 electrode at $1M-LiPF_6/PC-DEC(1:1)$ was small leak current of 0.0004A for 15min, long voltage retention of 0.8V after 100h and linear time-voltage curves with small IR-drop.

A Subgroup IB Isolate of Cucumber mosaic virus Isolated from Lagenaria leucantha var. gourda (표주박(Lagenaria leucantha var. gourda)에서 분리한 서브그룹 IB계통의 Cucumber mosaic virus)

  • Oh, Sun-Mi;Hong, Jin-Sung;Ryu, Ki-Hyun;Lee, Gung-Pyo;Choi, Jang-Kyung
    • Research in Plant Disease
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    • v.15 no.3
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    • pp.254-258
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    • 2009
  • An isolate of Cucumber mosaic virus (CMV), called as Lag-CMV, was identified from Lagenaria leucantha var. gourda showing mosaic symptom, and its properties was compared to Fny-CMV (subgroup IA) and As-CMV (subgroup IB) by host reaction in several indicator plants, dsRNA analysis, RT-PCR analysis, restriction enzyme profile of the PCR products and nucleotide sequence of coat protein gene. Lag-CMV was similar to As-CMV used as a control CMV by the induced chlorotic spot on inoculated leaves and mosaic symptoms on upper leaves of N. tabacum. cv. Xanthi nc. In the cucumber and zucchini squash, Lag-CMV and As-CMV induced a mild mosaic symptoms than that of Fny-CMV. Size and shapes of local lesions on Chenophodium amaranticolor and Vigna unguiculata induced by Lag-CMV was similar those by Fny-CMV or As-CMV. In experiments of dsRNA profiles and RT-PCR analysis of coat protein gene, Lag-CMV was come within subgroup I CMV. Moreover, restriction enzyme analysis using EcoRI, SalI, MspI, XhoI, and HindIII of the RTPCR products and nucleotide sequence analysis of the coat protein gene showed that Lag-CMV belong to a member of CMV subgroup IB of the same to As-CMV.

Monitoring of Blood Cytokines by PIT-1 Genotypes in Day 150 Male Pigs

  • Yoon, J.;Rhee, M.S.;Seo, D.S.;Kim, B.C.;Ko, Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.12
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    • pp.1659-1664
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    • 2001
  • Several studies have been done regarding carcass traits and growth in pigs. Recently, these have progressed to examine increases in economic traits, including meat quality and meat quantity, by using candidate genes. One of them is the pituitary-specific protein PIT-1, a member of the POU (Pit-Oct-Unc) family of transcription factors playing an important regulatory role in developmental processes. In addition, muscle development is known to be regulated in part by growth factors and cytokines locally produced. Therefore, studies were performed to analyze PIT-1 genotypes and serum cytokines (IGF-I, IGF-II, TGF-${\beta}1$, EGF, cortisol, DHEA-S, IL-2, and IL-6) in castrated male pigs for their possible involvement in the development of carcass traits. The genotypes of PIT-1 gene were analyzed by PCR-RFLP with MspI restriction enzyme. But, only CD and DD genotypes, not CC genotype, have been detected. Based on PIT-1 genotyping, a significant difference in EGF expression beween CD type (78.8 ng/ml) and DD type (46.0 ng/ml) was detected (p<0.05), whereas other cytokines did not show any statistical significance depending on PIT-1 genotypes. Collectively, these results suggest the possibility that EGF could affect the formation of carcass traits.

The Diversity of Culturable Organotrophic Bacteria from Local Solar Salterns

  • Yeon, Sun-Hee;Jeong, Won-Jin;Park, Jin-Sook
    • Journal of Microbiology
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    • v.43 no.1
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    • pp.1-10
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    • 2005
  • We isolated and cultured bacteria inhabiting solar saltern ponds in Taean-Gun, Chungnam Province, Korea. All of the isolated 64 strains were found to be moderately halophilic bacteria, growing in a salt range of 2-20 %, with an optimal concentration of 5% salt. Bacterial diversity among the isolated halophiles was evaluated via RFLP analyses of PCR-amplified 16S rDNAs, followed by phylogenetic analysis of the partial 16S rDNA sequences. The combination of restriction enzyme digestions with HaeIII, CfoI, MspI and RsaI generated 54 distinct patterns. A neighbor-joining tree of the partial 16S rDNA sequences resulted in the division of the 64 strains into 2 major groups, 45 strains of ${\gamma}-Proteobacteria$ (70.3%) and 19 strains of Firmicutes (29.7%). The ${\alpha}-Proteobacteria$ and Cytophaga-Flavobacterium-Bacterioides groups, which were repeatedly found to exist in thalassohaline environments, were not represented in our isolates. The ${\gamma}-Proteobacteria$ group consisted of several subgroups of the Vibrionaceae (37.5%), Pseudoalteromonadaceae (10.9%), Halomonadaceae (7.8%), Alteromonadaceae (7.8%), and Idiomarinaceae (6.3%). Members of Salinivibrio costicola (29.7%) were the most predominant species among all of the isolates, followed by Halobacillus treperi (12.5%). Additionally, three new species candidates were found, based on similarities of the 16S rDNA sequences to those of previously published species.

Hypermethylation Status of E-Cadherin Gene in Gastric Cancer Patients in a High Incidence Area

  • Rashid, Haroon;Alam, Khursheed;Afroze, Dil;Yousuf, Adfar;Banday, Manzoor;Kawoosa, Fizalah
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.6
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    • pp.2757-2760
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    • 2016
  • Gastric cancer (GC) is the fourth most prevalant cancer and the second leading cause of cancer-related mortality worldwide. As in other cancers gastric carcinogenesis is multifactorial involving environmental, genetic and epigenetic components. Epigenetic silencing due to hypermethylation of tumour suppressor genes is one of the key events in gastric carcinogenesis. This study was aimed to analyse the hypermethylation status of the E-Cadherin (CDH1) gene promoter in GCs in the ethnic Kashmiri population. In this study a total of 80 GC patients were recruited. Hypermethylation in tumour tissue was detected by methylation specific PCR (MS-PCR). Hypermethylation of CDH1 promoter was observed in 52 (65%) of gastric carcinoma cases which was significantly much higher than adjacent normal tissue [$p{\leq}0.0001$]. Further the frequency of CDH1 promoter methylation was significantly different with intestinal and diffuse types of gastric cancer [55.7% vs 82.1%; p<0.05]. Moreover females and cases with lymph node invasion had higher frequencies of CDH1 hypermethylation [$P{\leq}0.05$]. Thus the current data indicate a vital role of epigenetic alteration of CDH1 in the causation and development of gastric cancer, particularly of diffuse type, in our population.