• Title/Summary/Keyword: merA

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Generation of a Specific Marker to Discriminate Bacillus anthracis from Other Bacteria of the Bacillus cereus Group

  • Kim, Hyoung-Tai;Seo, Gwi-Moon;Jung, Kyoung-Hwa;Kim, Seong-Joo;Kim, Jee-Cheon;Oh, Kwang-Geun;Koo, Bon-Sung;Chai, Young-Gyu
    • Journal of Microbiology and Biotechnology
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    • v.17 no.5
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    • pp.806-811
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    • 2007
  • Bacillus anthracis is a soil pathogen capable of causing anthrax that is closely related to several environmental species, including B. cereus, B. mycoides, and B. thuringiensis. DNA homology studies showed that B. anthracis, B. cereus, B. mycoides, and B. thuringiensis are closely related, with a high sequence homology. To establish a method to specifically detect B. anthracis in situations such as environmental contamination, we initially performed RAPD-PCR with a 10-mer random primer and confirmed the presence of specific PCR bands only in B. anthracis species. One region specific for B. anthracis was cloned and sequenced, and an internal primer set was designed to amplify a 241-bp DNA fragment within the sequenced region. The PCR system involving these specific primer sets has practical applications. Using lyses methods to prepare the samples for PCR, it was possible to quickly amplify the 241-bp DNA segment from samples containing only a few bacteria. Thus, the PCR detection method developed in this study is expected to facilitate the monitoring of environmental B. anthracis contamination.

Purification and Enzymatic Properties of Myrosinase in Korean Mustard Seed(Brassica juncea) (한국산 겨자중 Myrosinase의 정제 및 효소학적 특성)

  • 신창식;서권일;강갑석;안철우;김용관;심기환
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.25 no.4
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    • pp.687-694
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    • 1996
  • Myrosinase was purified from Korean mustard seed(Brassica juncea) by a sequential process of DEAE-cellulose, concanavalin A-sepharose, and Superose 6 chromatography. The molecular weight of puri-fied myrosinase(II-2) determined by SDS-polyacrylamide electrophoresis was 67KD. About a 248-fold purification for myrosinase II-2 was obtained after Superose 6 chromatography. Optimum pH of the myrosinase was 7.0 and optimum temperature of the enzyme was $3^{\circ}C.$ The enzyme was stable at pH 7.0, and below $30^{\circ}C.$ Cu, Hg and Fe ion significantly inhibited the enzyme activity, but ascorbic acid enhanced, resulting in a maximum activity by 1mM ascorbic acid. Among tile ascorbic acid ana-logues, dehydroascorbic acid inhibited the enzyme activity, whereas others showed a little effect. Reducing agents such as 2-mercaptoethanol and dithiothreitol inhibited the enzyme activity, but the reducing agents with ascorbic acid was enhanced enzyme activity.

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Genetic Diversity Based on Morphology and RAPD Analysis in Vegetable Soybean

  • Srinives, P.;Chowdhury, A.K.;Tongpamnak, P.;Saksoong, P.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.46 no.2
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    • pp.112-120
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    • 2001
  • Genetic diversity of 47 East-Asian vegetable soybean was characterized by means of agro-morphological traits and RAPD markers. A field trial was conducted to evaluate 14 agro-morphological traits. To study RAPD-based DNA analysis, a total of sixty 10-mer random primers were screened. Of these, 23 polymorphic markers in 16 varieties used for screening. Among 207 markers amplified, 48 were polymorphic for at least one pairwise comparison within the 47 varieties. A higher differentiation level between varieties was observed by using RAPD markers compared to morphological markers. Correspondence analysis using both types of marker showed that RAPD data could fully discriminate between all varieties, whereas morphological markers could not achieve a complete discrimination. Genetic distances between the varieties were estimated from simple matching coefficients, ranged from 0.0 to 0.640 with an average of 0.295$\pm$0.131 for morphological traits and 0.042 to 0.625 with an average of 0.336$\pm$0.099 for RAPD data, respectively. Cluster analysis based on genetic dissimilarity of these varieties gave rise to 4 distinct groups. The clustering results based on RAPDs did not match with those based on morphological traits. Geographical distribution of most varieties in each of the groups were not well defined. The results suggested that the level of genetic diversity within this group of East-Asian vegetable soybean varieties was sufficient for a breeding program and can be used to establish genetic relationships among them with unknown or unrelated pedigrees.

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DOWN REGULATION OF TGF-$\beta$ GENE EXPRESSION BY ANTISENSE OLIGO-DEOXYNUCLEOTIDES INCREASE rIFN-${\gamma}$-INDUCED NITRIC OXIDE SYNTHESIS IN MURINE PERITONEAL MACROPHAGES

  • Jun, Chang-Duk;Kim, Su-Ung;Lee, Seong-Yong;Chung, Hun-Taeg
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1995.04a
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    • pp.78-78
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    • 1995
  • Increasing evidence indicates that the production of nitric oxide (NO) by inducible NO synthase (NOS) is tightely regulated. Transforming growth factor-${\beta}$ (TGF-${\beta}$) is a homodimeric protein secreted during macrophage activation, but several lines of evidence suggest that TGF-${\beta}$ is selectively suppressive for macrophage NO production. We therefore reasoned that a strategy employing oligodeoxynucleotides(ODNs) complemently to TGF-${\beta}$ mRNA (antisense ODNs) might increase NO production in IFN-${\gamma}$-treated murine peritoneal macrophages. To evaluate this concept, we tested the effects of antisense ODNs targeted to TGF-${\beta}$ mRNA (25-mer ODNs complemently to TGF-${\beta}$mRNA sequences) by introducing it into the medium of cultured macrophages. Phosphorothiolation of ODNs were employed to retard their degradation. Antisense ODNs had no effect on NO production by itself, whereas IFN-${\gamma}$ alone had modest effect. When antisense ODNs were used in combination with IFN-${\gamma}$, there was a marked cooperative induction of NO production, These effects of antisense ODNs were associated with decreased TGF-${\beta}$ expression in activated macrophages. ODNs with the same nucleotides but a scrambled sequence had no effect. Adding anti-TGF-${\beta}$ antibodies to the IFN-${\gamma}$-treated macrophages mimicked the positive effect of antisense ODNs on NO production. In addition, the effects of either antisense ODNs or anti-TGF-${\beta}$ antibodies were blocked by adding TGF-${\beta}$ in cultured macrophages. These results indicate that the generation of TGF-${\beta}$ by activated macrophages provides a self-regulating mechanism by which the temporal and perhaps spatial production of NO, a reactive and potentially toxic mediator, can be finely regulated.

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TISSUE HEALING RESPONSE OF INCISED WOUND SUTURED BY STAPLES AND VARIOUS SUTURE MATERIALS (Staple과 수종의 봉합물의 봉합부위 창상치유 조직반응)

  • Suh, Min-Jung;Lee, Jae-Hun
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.18 no.3
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    • pp.500-514
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    • 1996
  • The purpose of this study was to observe the tissue response in applying staples and various suture materials to both scalp and buccal mucosa in rabbits. 18 rabbits were divided into 6 groups. The incised wounds of both scalp and buccal mucosa were sutured with staples, polyglactin 910, chromic catgut, mer silk and nylon. The experimental animals were sacrificed after 1, 3, 5, 7, 10, 14 days posto peratively 3 animals at one time. The tissue was stained with Hematoxylin and eosin, and Masson's Trichrom. In light microscopic examinations, the sutured sites were examined histologi cally according to 6 degrees about inflammation and collagen deposit. The results were obtained as follows, 1. The chromic catgut, an absorbable suture material, was absorbed by 7 days, whereas polyglactin 910 and mersilk began to get absorbed after 7 days. 2. Mersilk manifested a broad range of inflammation in the scalp, and both staple and nylon showed a severe inflammatory reaction in the buccal mucosa. 3. With polyglactin 910, both tissue samples showed only minor foreign body reaction, however in the scalp, the process of fibrosis took place compara tively slowly, whereas in the buccal mucosa, it occurred promptly and manifested active fibrosis by 7 days. 4. Mersilk showed widespread a matrix formation in both scalp and buccal mucosa, and showed the most severe inflammatory reaction by 3 days, which did not seem to decrease even after 7 days. 5. Both staple and nylon showed relatively a severe inflammatory reaction, however fibrosis took place rather promptly compared to the other groups. 6. Generally, in the buccal mucosa fibrosis occurred more promptly than in the scalp in both control and experimental groups. 7. Retention of the suture material and stability of the knot were the best with the staple, and better stability was manifested by the multi-stranded poly glactin 910 and mersilk than singlestranded chromic catgut and nylon. From above results, in the buccal mucosa absorbable suture materials especially polyglactin 910 showed better response in the aspect of inflammatory reaction, while in the scalp monofilament suture materials such as staple and nylon manifested a early fibrosis and collagen formation.

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Identification of Molecular Markers Linked to Lf2 Locus in Soybean

  • Kim Myung-Sik;Park Min-Jung;Jeong Woo-Hyeun;Nam Ki-Chul;Chung Jong-Il
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.2
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    • pp.169-172
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    • 2006
  • Leaflet number of soybean controlled by Lf2 locus is the important trait in photosynthesis and plant type. The objective of this research was to identity molecular markers linked to the lf2 locus. A total of $115F_2$ plants were derived from a cross between normal three-leaflet type Sinpaldalkong (Lf2Lf2) and seven-leaflet mutant type T255 (lf2lf2). All leaflet counts of parents and $F_2$ individual plants were made in the field on fully expanded leaves on the main stem when terminal growth of the main stem had ceased. One-thousand 10-mer oligonucleotide RAPD primers and 664 SSR primers were used. The segregation ratios of 3 : 1 were observed in the $F_2$ population and the Chi-square values strongly suggested that the seven-leaflet was controlled by a single recessive gene. A genetic map was constructed from the 15 segregating markers (9 RAPDs, 5 SSRs, 1 lf2 locus). OPAD03 and OPAI13 RAPD markers were linked to the lf2 locus that controlled seven-leaflet type at a distance of 20.5 and 23.5 cM, respectively. Molecular markers identified in this study linked with lf2 locus will be helpful to locate lf2 locus on the public soybean molecular linkage map and would be useful for tagging the lf2 locus that controls seven-leaflet trait.

Biological, Physical and Cytological Properties of Pepper mottle virus-SNU1 and Its RT-PCR Detection

  • Han, Jung-Heon;Choi, Hong-Soo;Kim, Dong-Hwan;Lee, Hung-Rul;Kim, Byung-Dong
    • The Plant Pathology Journal
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    • v.22 no.2
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    • pp.155-160
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    • 2006
  • A strain of Pepper mottle virus (PepMov) was isolated from chili pepper plants in Korea. In host range study, this virus, designated PepMoV-SNU1, shared most characteristics with PepMoV isolates reported previously. Thermal inactivation point ($45^{\circ}C\;to\;75^{\circ}C$) and dilution end point ($10^{-1}\;to\;10^{-4}$) of PepMoV-SNU1 showed differences depending on the propagation hosts. Cylindrical and pinwheel-shaped inclusions were always observed in pepper leaf tissues infected with the virus alone. Unexpectedly, a special structure of pinwheel shaped inclusion surrounded with unknown small spots was also observed in the leaf section when co-infected with a strain of pepper mild mottle virus. The partial sequence of coat protein gene and 3' untranslated region of PepMoV-SNU1 showed 98% identity with those of other PepMoV isolates. A primer pair derived from 3' end of the coat protein gene and poly A tail regions were designed. Optimal detection condition of PepMoV-SNU1 by RT-PCR was tested to determine appropriate annealing temperature and additional volumes of oligo-dT (18-mer), dNTP, and Taq polymerase. Under the optimized condition, an expected 500 Up PCR-product was detected in pepper leaves infected with PepMoV-SNU1 but not in healthy plants.

A RFID Tag Indexing Scheme Using Spatial Index (공간색인을 이용한 RFID 태그관리 기법)

  • Joo, Heon-Sik
    • Journal of the Korea Society of Computer and Information
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    • v.14 no.7
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    • pp.89-95
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    • 2009
  • This paper proposes a tag indexing scheme for RFID tag using spatial index. The tag being used for the inventory management and the tag's location is determined by the position of readers. Therefore, the reader recognizes the tag, which is attached products and thereby their positions can be traced down. In this paper, we propose hTag-tree( Hybrid Tag index) which manages RFID tag attached products. hTag-tree is a new index, which is based on tag's attributes with fast searching, and this tag index manages RFID tags using reader's location. This tag index accesses rapidly to tags for insertion, deletion and updating in dynamic environment. This can minimize the number of node accesses in tag searching comparing to previous techniques. Also, by the extension of MER in present tag index, it is helpful to stop the lowering of capacity which can be caused by parent node approach. The proposed index experiment deals with the comparison of tag index. Fixed Interval R-tree, and present spatial index, R-tree comparison. As a result, the amount of searching time is significantly shortened through hTag-tree node access in data search. This shows that the use of proposed index improves the capacity of effective management of a large amount of RFID tag.

Stability of Human Centromeric Alphoid DNA Repeat during Propagation in Recombination-Deficient Yeast Strains (효모의 재조합 변이주를 이용한 인간 Centromeric Alphoid DNA Repeat의 안정성에 관한 연구)

  • Kim, Kwang-Sup;Shin, Young-Sun;Lee, Sang-Yeop;Ahn, Eun-Kyung;Do, Eun-Ju;Park, In-Ho;Leem, Sun-Hee;SunWoo, Yang-Il
    • Korean Journal of Microbiology
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    • v.43 no.4
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    • pp.243-249
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    • 2007
  • The centromere is a highly differentiated structure of the chromosome that fulfills a multitude of essential mitotic and meiotic functions. Alphoid DNA (${\alpha}$-satellite) is the most abundant family of repeated DNA found at the centromere of all human chromosomes, and chromosomes of primates in general. The most important parts in the development of Human Artificial Chromosomes (HACs), are the isolation and maintenance of stability of centromeric region. For isolation of this region, we could use the targeting hook with alphoid DNA repeat and cloned by Transformation-Associated Recombination (TAR) cloning technique in yeast Saccharomyces cerevisiae. The method includes rolling-circle amplification (RCA) of repeats in vitro to 5 kb-length and elongation of the RCA products by homologous recombination in yeast. Four types of $35\;kb{\sim}50\;kb$ of centromeric DNA repeat arrays (2, 4, 5, 6 mer) are used to examine the stability of repeats in homologous recombination mutant strains (rad51, rad52, and rad54). Following the transformation into wild type, rad51 and rad54 mutant strains, there were frequent changes in inserted size. A rad52 mutant strain showed extremely low transformation frequency, but increased stability of centromeric DNA repeat arrays at least 3 times higher than other strains. Based on these results, the incidence of large mutations could be reduced using a rad52 mutant strain in maintenance of centromeric DNA repeat arrays. This genetic method may use more general application in the maintenance of tandem repeats in construction of HAC.

Genetic Variability Based on Randomly Amplified Polymorphic DNA in Kacip Fatimah (Labisia pumila Benth & Hook f) collected from Melaka and Negeri Sembilan States of Malaysia

  • Bhore, Subhash J.;Nurul, A.H.;Shah, Farida H.
    • Journal of Forest and Environmental Science
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    • v.25 no.2
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    • pp.93-100
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    • 2009
  • In Malaysia, Labisia pumila Benth & Hook f, popularly known as 'Kacip Fatimah' has been used traditionally to treat various elements of the woman's health in Malay community. The objective of this study was to develop randomly amplified polymorphic DNA (RAPD) based DNA markers for the identification of L. pumila and to distinguish its three varieties from each other. Total DNA from nine accessions of L. pumila was extracted by CTAB method and polymerase chain reactions (PCR) were carried out to amplify the segments of DNA using different primers to develop DNA barcode using RAPD technique. To find out variety-specific DNA marker/s, twenty different 10-mer primer sequences with annealing temperature from 36-$40^{\circ}C$ were evaluated in triplicate. Out of 20 random primers, two primers (OPA-1 and OPA-2/A10) were selected which produced reliable RAPD band patterns. To have DNA based handle, two RAPD amplification products were cloned and sequenced to determine the identity of the DNA. RAPD analysis using two random primers generated 72 discrete bands ranging in size 200 bp-3,000 bp. Fifty nine of these were polymorphic loci (82%) and thirteen were non-polymorphic loci (18%). A total of 32 bands polymorphic loci (72%) were amplified with primer OPA-1 and analyzed by cluster analysis and UPGMA (Unweighted Pair Group Method with Arithmetic) to present a dendogram depicting the degree of genetic relationship among nine accessions of L. pumila. Our results shows the reasonable genetic diversity among the L. pumila varieties and within varieties; and two RAPD marker sequences obtained could be used to identify L. pumila at species level.

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