• Title/Summary/Keyword: Restriction map

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HiCORE: Hi-C Analysis for Identification of Core Chromatin Looping Regions with Higher Resolution

  • Lee, Hongwoo;Seo, Pil Joon
    • Molecules and Cells
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    • v.44 no.12
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    • pp.883-892
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    • 2021
  • Genome-wide chromosome conformation capture (3C)-based high-throughput sequencing (Hi-C) has enabled identification of genome-wide chromatin loops. Because the Hi-C map with restriction fragment resolution is intrinsically associated with sparsity and stochastic noise, Hi-C data are usually binned at particular intervals; however, the binning method has limited reliability, especially at high resolution. Here, we describe a new method called HiCORE, which provides simple pipelines and algorithms to overcome the limitations of single-layered binning and predict core chromatin regions with three-dimensional physical interactions. In this approach, multiple layers of binning with slightly shifted genome coverage are generated, and interacting bins at each layer are integrated to infer narrower regions of chromatin interactions. HiCORE predicts chromatin looping regions with higher resolution, both in human and Arabidopsis genomes, and contributes to the identification of the precise positions of potential genomic elements in an unbiased manner.

Development of Restriction Fragment Length Polymorphism(RELP) Markers in Silkworm, Bombyx mori (누에 RFLP(제한단편 다형현상)마커 개발)

  • 고승주;김태산;이영승;황재삼;이상몽
    • Korean journal of applied entomology
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    • v.36 no.1
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    • pp.96-104
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    • 1997
  • A silkworm Bombyx mori genomic DNA library was constructed from polyphagous J111 strain and unpolyphagous $C_3$ strain to develop the genomic study by DNA makers. Genomic DNAs of two strains were digested with restriction enzyme EcoRI and ligated into pUC18. The ligated plasmids were transferred into E. coli host strain DH5$\alpha$. When the genomic DNAs were hybridized with insert DNAs from transformant, could be categorized from hybridization patterns to three groups as high repetitive sequence, moderately repetitive sequence, and low-copy number sequences. A total of 219 clones containing single or low-copy number sequence inserts were examined for any polymorphisms between two strains of J111 and $C_3$. Forty six clones showed RFLPs and 10 of these clones were used as a probe of analysis of $F_2$ population derived from crossing between J111 and $C_3$ strain. The genetic inheritance tested with each clones will be important tools to construct the genetic map of the silkworm, Bombyx mori.

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A Case Study on the Status and Problems of Regulations of Land Use in Gyeonggi-do (경기도 토지이용규제 현황과 문제점 고찰)

  • Kim, Young-Hoon;Kwon, Gyoung-Nam
    • KIEAE Journal
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    • v.16 no.6
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    • pp.57-67
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    • 2016
  • Purpose: Laws and regulations of land use are enormous, and the appliance of regulations is overlapped redundantly. Therefore, there are many problems such as time consuming in the process, limiting individual property rights, and interrupting enterprises' economic activities. This study will discuss problems of redundant regulations of land use and its improvement by figuring out current regulations of land use in Gyeonggi-do, one of provinces which applies the most various regulations of land use. Method: This study reviews laws on national land-use planning system and characteristics of land-use regulation in Korea. The extent of the review is limited to "framework act on the regulation of land use" with categories of national land, urban planing, architecture, etc. Through case studies in Gyeonggi-do, the status and problems of redundant regulations of land use are defined. For example, it is overlapped in "Seoul Metropolitan Area Readjustment Planning Act", Development Restriction Zone, Paldang Special District, and so on. It is mainly referred to 2015 Gyeonggi-do land-use restriction map. Result: First, Gyeonggi-do confronts many problems related to the development restrictions and the financial increasement for environmental management by redundant regulations. The development restrictions include supplying additional land for industrial use, relocating colleges, and height limitation relating to military facilities. Second, in order to organize redundant regulations, it is required to combine similar regulations and adjust through communication system among other departments. Third, regulations should consider unique local condition of each district. Lastly, efficient application of regulations is necessary so as to maximize the function of land, protect individual property rights, and stimulate local development.

Studies on KEM1 Gene Controlling Mitotic Cell Division in Yeast: Molecular Cloning of a High Copy Suppressor (ROK1) of kem1 (효모에서 세포분열을 조절하는 KEM1 유전자에 관한 연구: kemi의 High Copy Suppressor (ROK1) 클로닝)

  • Kim, Sang Hyeon;Kim, Jin Mi
    • Korean Journal of Microbiology
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    • v.30 no.1
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    • pp.37-41
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    • 1992
  • The KEM1 gene is known to affect microtubule and spindle pole body function during the cell division cycle in Saccharomjyces cerevisiae. To identify new genes with functions similar or related to those of KEM1, we isolated a high copy suppressor gene (ROK1) that suppresses the kem1 mutation when cloned on a high copy number plasmid but not on a low copy number plasmid. Two clones which suppress both the benomyl hypersensitivity and the $Kar^{-}$ enhancing phenotype of kem1 null mutation were isolated and were shown to have a 9.0 kb identical insert by restriction endonuclease analysis. The restriction map constructed indicates that this suppressor gene, ROK1 is not KEM1. Subcloning experiments suggest that the functional region of ROK1 is at least 3.0kb in size.

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Molecular Cloning of nod Genes from Bradyrhizobium sp. SNU001 (Bradyrhizobium sp. SNU001 nod 유전자 클로닝)

  • 고세리;심웅섭;안정선
    • Korean Journal of Microbiology
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    • v.30 no.4
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    • pp.246-251
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    • 1992
  • Molccular cloning of nod genes from Bradvrhizobium sp. SNU001, a nitrogen-fixing symbiont isolated from thc root nodules of soybean (Clycine trim) . was carried out. nod genes were found to be located on thc genome of the symbiont by gcnomic hybridization with 4.5 kb EcoRI/HndIII fragment (nod DABC) of Rhizohium meliloti as probe. Genomic library of this symbiont was constructed using h phage EMBL3-BanlHI vector. from which five nod positive clones were sclectcd by primary and secondary screening methods. The partial restriction map of inserted genomic DNA of h CNS-l(c1one 2) was constructed. and 3.9 kh Bun7HI fragment. which showed strong hybridization signal to the probe, was subcloned into pBS KS(+) plasmid vector. Partial restriction inap ot' a selected subclone (pBjCNS-I) was constructed and nod DABC was found to be located on the 1.8 kb KpnI/Sacl fragment of this subclone.

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Molecular Cloning of the Arginine Biosynthetic Genes from Corynebacterium glutamicum

  • Chun, Jae-Shick;Jung, Sam-Il;Ko, Soon-Young;Park, Mee-Young;Kim, Soo-Young;Lee, Heung-Shick;Cheon, Choong-Ill;Min, Kyung-Hee;Lee, Myeong-Sok
    • Journal of Microbiology
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    • v.34 no.4
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    • pp.355-362
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    • 1996
  • Complementation cloning of the argC, E, B, D, F, and G genes in Corynebacterium glutamicum was done by transforming the genomic DNA library into the corresponding arginine auxotrophs fo Escherichia coli. Recombinant plasmids containing 6.7 kb and 4.8kb fragments complementing the E. coli argB mutant were also able to complement the E. coli argC, E, A, D, and F mutants, indicating the clustered organization of the arginine biosynthetic genes within the cloned DNA fragments. The insert DNA fragments in the recombinant plasmids, named pRB1 AND pRB2, were physically mapped with several restriction enzymes. By further subcloning the entire DNA fragment containing the functions and by complementation analysis, we located the arg genes in the order of ACEBDF on the restriction map. We also determined the DNA nucleotide sequence of the fragment and report here the sequence of the argB gene. When compared to that with the mutant strain, higher enzyme activity of N-acetylglutamate kinase was detected in the extract of the mutant carrying the plasmid containing the putative argB gene, indicating that the plasmid contains a functional argB gene. Deduced amino acid sequence of the argB gene shows 45%, 38%, and 25% identity to that from Bacillus strearothermophilus, Bacillus substilus, and E. coli respectively. Our long term goal is genetically engineering C. glutamicum which produces more arginine than a wild type strain does.

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Molecular Cloning and Expression of Bacillus stearothermophilus $\beta$-D-Xylosidase Gene in E. coli (Bacillus stearothermophilus $\beta$-D-Xylosidase 유전자의 크로닝 및 Escherichia coli에서의 발현)

  • 오세욱;박성수;최용진;박영인
    • Microbiology and Biotechnology Letters
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    • v.20 no.2
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    • pp.136-142
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    • 1992
  • Bacillus stearothemophilus isolated from soil was identified to express multiple extracellular xylanases. Two HindIII restriction fragments of 5.4 and 6.4 kb from B. stearothermophilus genomic DNA were cloned into pBR322 to obtain recombinant plasmids pMG0l and pMG02, respectively, which enabled E. coli HBlOl cells to produce $\beta$-D-xylosidase activity. By subcloning into pUC18 and Southern blotting, the loci of the $\beta$-D-xyiosidase genes were elucidated to be on non-homologous DNA fragments of 2.2 kb from pMGOl(pMG1) and 1.0 kb from pMG02(pMG2), respectively. The two enzymes produced in E. coli cleaved xylobiose, xylotriose, xylotetrose and xylotetrose to produce xylose as a major end product. The gene on pMG1, distinct from that on pMG2 was observed to encode a bifunctional protein that displayed both P-D-xylosidase (EC.3.2.1.37) and a-L-arabinofuranosidase activities (EC.3.2.1.55).

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Improvement in Legislative Assessment of the Environmental Conservation Value Assessment Map Considering the Restriction on Acts of Special-Purpose Areas (용도 지역 행위 제한을 고려한 국토환경성평가지도 법제적 평가 개선)

  • Yang, Hyun-Jae;Kim, Geun-Han;Yoon, Jeong-Ho;Jun, Chul-Min;Lee, Eun-Jung;Hwang, So-Young
    • Journal of the Korean Society of Environmental Restoration Technology
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    • v.21 no.1
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    • pp.13-30
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    • 2018
  • In terms of environmental friendly land use and objective environmental assessment, legislative assessment items of Environmental Conservation Value Assessment Map (ECVAM) consists of designated areas related to conservation and protection, and areas that are planned to be designated in the future. However, the gap with the reality due to omission of several protection areas and land use regulations, and assessment grades according to 3 division land use map of forest, agricultural land, and urbanized area, results in low application and utilization of ECVAM. Therefore in this study, the legislative assessment of the ECVAM was performed with new assessment items and its new ratings, to suggest an improvement in legislative assessment items of the ECVAM. As a result, legally protected areas of inhabited islands under absolute conservation, special wildlife protection districts, protected marine areas, environmental preservation sea areas, scenic spots, forest protection zone, traditional temple preservation zone, and 45 zone or district related to regulation of land use were additionally designated as new legislative assessment items. New grade ratings were given to each additional assessment items in consideration of the restrictions on acts. As a result of the legislative assessment based on the new assessment items and new grades, the 1st grade area increased by 3.47%, and the 2nd grade area increased by 19.35%. The 3rd grade area decreased by 8.54%, the 4th grade area increased by 2.95%, and 5th grade increased by 2.91%. In addition, the out-of-grade area decreased by 20.14%, considered to be a realistic assessment based on land use. With the improved legislative assessment, it is possible to provide a more accurate environmental assessment map. Increase usage of ECVAM is expected in providing regulations of land use and base data for integrated land management of land environmental planning.

Construction of the Novel Baculovirus Transfer Vector Using the p10 Gene of BmNPV (BmNPV의 p10 유전자를 이용한 새로운 전이벡터 개발)

  • 강석우;진병래
    • Journal of Sericultural and Entomological Science
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    • v.39 no.2
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    • pp.180-185
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    • 1997
  • To develope the novel baculovirus transfer vector, the p10 gene was cloned from the Bombyx mori nuclear polygedrosis virus (BmNPV) vB2 strain isolated from the B. mori larvae of sericultural farms. The novel transfer vector was constructed by using the p10 gene of BmNPV vB2 strain was 210 bp. The TAAG sequence at the -71 bp of upstream from translation initiator ATG and two polyadenylation signal site at the downstream from terminator TAA were also detected in the p10 gene. The 5' and 3' flanking region of the p10 gene amplified by PCR was cloned into pBluescriptII SK(+) and then transfer vector pBm10 was construceted. The 7.9 kb pBm10 was analysed by restriction enzymes and the map was confirmed. In order to determine the expression of foreign gene of pBm10, $\beta$-galactosidase gene was inserted in the SmaI site of foreign gene cloning site of pBm10. The pBm10 containing $\beta$-galactosidase gene was cotranfected wth genomic DNA of BmNPV vB2 into BmN-4 cells. The recombinant baculovirus expressing $\beta$-galactosidase was also produced polygedra in the infected cells. The results indicated that pBm10 is functional, suggesting that in the baculovirus expression vector system, the recombinant virus produced by pBm10 was effective by oral infection for the producing recombinant proteins in in vivo expression.

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Characterization of Recombinant Derivatives of pJY711 of Multicopy Streptomyces Plasmid (Multicopy Streptomyces 플라스미드 pJY711의 재조합 유도체의 특성)

  • 염도영;공인수;유주현
    • Korean Journal of Microbiology
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    • v.28 no.1
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    • pp.35-40
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    • 1990
  • The restriction clevage map of multi-copy recombinant plasmid, pJY712(8.1kb), carrying the thiostrepton resistance gene(tsr) was determined. pJY712 had a broad host range in Streptomyces and contained single BglII site for cloning purpose. The plasmid showed the phenomenon of lethal zygosis ($Ltz^{+}$). Transformation frequency of pJY712 was $5.0\times 10^{4}$ transformants per ug plasmid DNA (TFU) in S. lividans. Plasmid pJY713 was constructed by inserting the tyrosinase gene(mel) into the BclI site of pJY712. Recombinant plasmid pJY714 carrying the mel gene was constructed by in vitro deletion of a segment (1.9kb BglII-BclI fragment) from pJY713.

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