• Title/Summary/Keyword: 유전체 분석

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Cytogenetic Study of Maackia amurensis Rupr. & Maxim. and M. fauriei (Levl.) Takeda Using Karyotyping Analysis and the FISH Technique (핵형분석과 FISH 기술을 이용한 솔비나무와 다릅나무의 세포유전학적 연구)

  • Kim, Soo-Young;Kim, Chan-Soo
    • Korean Journal of Plant Taxonomy
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    • v.39 no.3
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    • pp.193-198
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    • 2009
  • Chromosome analysis using karyotyping and bicolor FISH were carried out for two Maackia species (M. fauriei and M. amurensis) found in Korea. The somatic metaphase chromosome number was 2n = 2x = 18 in both, and the size of these chromosomes ranged from 3.58 to $5.82{\mu}m$. The chromosome complements consisted of two pairs of metacentric (chromosomes 1 and 7), four pairs of submetacentrics (chromosomes 4, 6, 8 and 9) and three pairs of subtelocentrics (chromosomes 2, 3 and 5) in M. fauriei but, chromosomes 4 (subtelocentric) and 7 (submetacentric) of M. amurensis have different morphology. Using bicolor FISH, a pair of 45S rDNA loci were observed for both M. fauriei and M. amurensis, but the number and site of the 5S rDNA signal were different in the two species. M. fauriei has two pairs of 5S signals on chromosomes 7 and 8 but, M. amurensis has four paris on chromosomes 3, 4, 7 and 7. Hence, the 5S rDNA is a useful FISH for Maackia species.

A Single Nucleotide Deletion resulting in Frameshift in Two Korean Neonates with Thyroxine-Binding Globulin Deficiency (단일 뉴클레오타이드 결손으로 인한 Frameshift 돌연변이로 규명된 티록신결합글로불린 결핍증 1례)

  • Park, Sang-Joon;Suh, Jin-Soon;Jung, Min-Ho;Lee, Hee-Jin;Suh, Byung-Kyu;Lee, Won-Bae;Lee, Byung-Churl
    • Clinical and Experimental Pediatrics
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    • v.48 no.11
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    • pp.1252-1255
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    • 2005
  • Abnormalities in the levels of thyroxine-binding globulin (TBG) are not associated with clinical disease and they do not require treatment. Congenital TBG deficiency is inherited in an X-linked manner. To date, some complete and partial TBG variants and one polymorphism have been identified by analysis of the TBG gene. Two male neonates were referred to us because of their low $T_4$ levels that were noted on the neonatal screening test. They showed normal levels of free $T_4$ and TSH. Their serum TBG was not detectable and those values of their parents were within the normal ranges. The genomic DNA was extracted from their white blood cells and the four coding exons of the TBG gene were amplified by using polymerase chain reaction. Sequencing of the four coding regions and all the intron/exon junctions revealed a single nucleotide deletion of the first base of the codon 352 of the mature protein in both of the neonates. This mutation resulted in a frameshift and a premature stop codon (TGA) 374. Their mothers were shown to be heterozygotes. We detected a single nucleotide deletion resulting in a frameshift in two male Korean neonates who had complete TBG deficiency.

A Case of Congenital Nephrogenic Diabetes Insipidus Confirmed by Gene Analysis (유전자 분석검사로 확진된 선천성 신성 요붕증 1례)

  • Cho, Eun Young;Oh, Jin Hee;Koh, Dae Kyun
    • Clinical and Experimental Pediatrics
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    • v.48 no.6
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    • pp.669-674
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    • 2005
  • Nephrogenic diabetes insipidus (NDI) is a disorder in which the secretion of antidiuretic hormone is normal, but the response of the renal collecting tubules to vasopressin is impaired. Compared with acquired NDI (a-NDI), which is secondary to chronic bilateral incomplete urinary tract obstruction with hydronephrosis, congenital NDI (c-NDI) is a very rare heritable disorder that usually follows the X- linked recessive pattern. Clinical symptoms of c-NDI can be non specific, and often the disease ultimately results in failure to thrive, or mental retardation. Recently, the diagnosis can be confirmed by direct sequencing analysis of the peripheral blood specimens. The long-term results of treatment for c-NDI are not satisfactory. Reports on the follow up of c-NDI cases are rare and there is no report on the cases treated with combinations of three drugs. We report herein a case of severe c-NDI in an 8 year-old-boy with a severely dysconfigurated urinary tract system. The patient and his mother showed a frameshift mutation on the AVPR2 gene on chromosome Xq28:.847_851delTGCTG (p.C283fsX90). The patient showed normal growth and development by treatment with combinations of hydrochlorothiazide ($65mg/m^2$), amiloride (0.3 mg/kg/d) and indomethacin ($100mg/m^2$), yet after five years he needed adjuvant cystostomy to relieve him from the residual symptoms of urgency with polyuria.

Cloning and Characterization of Xylanase 11B Gene from Paenibacillus woosongensis (Paenibacillus woosongensis의 Xylanase 11B 유전자 클로닝과 특성분석)

  • Yoon, Ki-Hong
    • Microbiology and Biotechnology Letters
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    • v.45 no.2
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    • pp.155-161
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    • 2017
  • A gene coding for the xylanase predicted from the partial genomic sequence of Paenibacillus woosongensis was cloned by PCR amplification and sequenced completely. This xylanase gene, designated xyn11B, consisted of 1,071 nucleotides encoding a polypeptide of 356 amino acid residues. Based on the deduced amino acid sequence, Xyn11B was identified to be a modular enzyme, including a single carbohydrate-binding module besides the catalytic domain, and was highly homologous to xylanases belonging to glycosyl hydrolase family 11. The SignalP4.1 server predicted a stretch of 26 residues in the N-terminus to be the signal peptide. Using DEAE-Sepharose and Phenyl-Sepharose column chromatography, Xyn11B was partially purified from the cell-free extract of recombinant Escherichia coli carrying a copy of the P. woosongensis xyn11B gene. The partially purified Xyn11B protein showed maximal activity at $50^{\circ}C$ and pH 6.5. The enzyme was more active on arabinoxylan than on oat spelt xylan and birchwood xylan, whereas it did not exhibit activity towards carboxymethylcellulose, mannan, and para-nitrophenyl-${\beta}$-xylopyranoside. The activity of Xyn11B was slightly increased by $Ca^{2+}$ and $Mg^{2+}$, but was significantly inhibited by $Cu^{2+}$, $Ni^{2+}$, $Fe^{3+}$, and $Mn^{2+}$, and completely inhibited by SDS.

DRAM Package Substrate Using Aluminum Anodization (알루미늄 양극산화를 사용한 DRAM 패키지 기판)

  • Kim, Moon-Jung
    • Journal of the Institute of Electronics Engineers of Korea SD
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    • v.47 no.4
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    • pp.69-74
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    • 2010
  • A new package substrate for dynamic random access memory(DRAM) devices has been developed using selective aluminum anodization. Unlike the conventional substrate structure commonly made by laminating epoxy-based core and copper clad, this substrate consists of bottom aluminum, middle anodic aluminum oxide and top copper. Anodization process on the aluminum substrate provides thick aluminum oxide used as a dielectric layer in the package substrate. Placing copper traces on the anodic aluminum oxide layer, the resulting two-layer metal structure is completed in the package substrate. Selective anodization process makes it possible to construct a fully filled via structure. Also, putting vias directly in the bonding pads and the ball pads in the substrate design, via in pad structure is applied in this work. These arrangement of via in pad and two-layer metal structure make routing easier and thus provide more design flexibility. In a substrate design, all signal lines are routed based on the transmission line scheme of finite-width coplanar waveguide or microstrip with a characteristic impedance of about $50{\Omega}$ for better signal transmission. The property and performance of anodic alumina based package substrate such as layer structure, design method, fabrication process and measurement characteristics are investigated in detail.

Genome-wide Copy Number Variation in a Korean Native Chicken Breed (한국 토종닭의 전장 유전체 복제수변이(CNV) 발굴)

  • Cho, Eun-Seok;Chung, Won-Hyong;Choi, Jung-Woo;Jang, Hyun-Jun;Park, Mi-Na;Kim, Namshin;Kim, Tae-Hun;Lee, Kyung-Tai
    • Korean Journal of Poultry Science
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    • v.41 no.4
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    • pp.305-311
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    • 2014
  • Copy number variation (CNV) is a form of structural variation that shows various numbers of copies in segments of the DNA. It has been shown to account for phenotypic variations in human diseases and agricultural production traits. Currently, most of chicken breeds in the poultry industry are based on European-origin breeds that have been mostly provided from several international breeding companies. Therefore, National Institute of Animal Science, RDA has been trying to restore and improve Korean native chicken breeds (12 lines of 5 breeds) for about 20 years. Thanks to the recent advance of sequencing technologies, genome-wide CNV can be accessed in the higher resolution throughout the genome of species of interest. However, there is no systematic study available to dissect the CNV in the native chicken breed in Korea. Here, we report genome-wide copy number variations identified from a genome of Korean native chicken (Line L) by comparing between the chicken reference sequence assembly (Gallus gallus) and a de novo sequencing assembly of the Korean native chicken (Line L). Throughout all twenty eight chicken autosomes, we identified a total of 501 CNVs; defined as gain and loss of duplication and deletion respectively. Furthermore, we performed gene ontology (GO) analysis for the putative CNVs using DAVID, leading to 68 GO terms clustered independently. Of the clustered GO terms, genes related to transcription and gene regulation were mainly detected. This study provides useful genomic resource to investigate potential biological implications of CNVs with traits of interest in the Korean native chicken.

Association of Single Nucleotide Polymorphism (SNP) in the PGK 2 Gene with Growth Traits in Pigs (돼지 PGK 2 유전자의 단일염기다형성 및 성장 형질과의 연관성 구명)

  • Jang, Hong-Chul;Kim, Sang-Wook;Lim, Da-Jeong;Kim, Jae-Young;Cho, Kyu-Ho;Kim, Myung-Jick;Lee, Ji-Woong;Choi, Bong-Hwan;Kim, Tae-Hun
    • Journal of Animal Science and Technology
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    • v.53 no.1
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    • pp.15-22
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    • 2011
  • The purpose of this study was to analyse of association between growth traits and single nucleotide polymorphisms (SNPs) polymorphism of phosphoglycerate kinase 2 (PGK 2) gene in pigs. The birth weight of piglet influences on weaning weight and survival rate that are import economic traits in pig industry. Also, these growth traits are representative factor to decrease a period getting to marketing weight as well as growth rate in pig. The PGK 2 is an isozyme that catalyzes the first ATP-generating step in the glycolytic pathwayand important enzyme related with energy metabolism. Twenty of SNPs were discoveredby genome structure analysis that compares the sequence on promoter and transcription region of PGK 2 gene in porcine chromosome 7. An association between PGK 2 SNPs and growth traits was analyzed in $F_2$ reciprocal-crossbred population between korean native pig (KNP) and Landrace. Association analysis indicated that polymorphism of the PGK 2 gene promoter region has significant effects on weight at birth (p<0.01) and weight at 3 weeks of age (p<0.0001). These results suggest that PGK 2 gene polymorphism was associated with energy metabolism and physiological function of growth in pig.

Novel substrate specificity of a thermostable β-glucosidase from the hyperthermophilic archaeon, Thermococcus pacificus P-4 (초고온 고세균 Thermococcus pacificus P-4로부터 내열성 β-glucosidase의 새로운 기질 특이성)

  • Kim, Yun Jae;Lee, Jae Eun;Lee, Hyun Sook;Kwon, Kae Kyoung;Kang, Sung Gyun;Lee, Jung-Hyun
    • Korean Journal of Microbiology
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    • v.51 no.1
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    • pp.68-74
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    • 2015
  • Based on the genomic analysis of Thermococcus pacificus P-4, we identified a putative GH1 ${\beta}$-glucosidase-encoding gene (Tpa-glu). The gene revealed a 1,464 bp encoding 487 amino acid residues, and the deduced amino acid residues exhibited 77% identity with Pyrococcus furiosus ${\beta}$-glucosidase (accession no. NP_577802). The gene was cloned and expressed in Escherichia coli system. The recombinant protein was purified by metal affinity chromatography and characterized. Tpa-Glu showed optimum activity at pH 7.5 and $75^{\circ}C$, and thermostability with a half life of 6 h at $90^{\circ}C$. Tpa-Glu exhibited hydrolyzing activity against various pNP-glycopyranosides, with kcat/Km values in the order of pNP-${\beta}$-glucopyranoside, pNP-${\beta}$-galactopyranoside, pNP-${\beta}$-mannopyranoside, and pNP-${\beta}$-xylopyranoside. In addition, the enzyme exhibited exo-hydrolyzing activity toward ${\beta}$-1,3-linked polysaccharide (laminarin) and ${\beta}$-1,3- and ${\beta}$-1,4-linked oligosaccharides. This is the first description of an enzyme from hyperthermophilic archaea that displays exo-hydrolyzing activity toward ${\beta}$-1,3-linked polysaccharides and could be applied in combination with ${\beta}$-1,3-endoglucanase for saccharification of laminarin.

Expression and Cloning of the pmmC Gene Encoding Phosphomannomutase in Sphingomonas chungbukensis DJ77 (Sphingomonas chungbukensis DJ77 균주에서 Phosphomannomutase를 암호화하는 pmmC 유전자의 클로닝과 발현)

  • Kim Mi-Hye;Choi Jung-Do;Shin Malshick;Kim Young-Chang
    • Microbiology and Biotechnology Letters
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    • v.33 no.2
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    • pp.84-89
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    • 2005
  • Phosphomannomutase (PMM) is a key enzyme in prokaryotes and eukaryotes, which catalyzes the conversion of ${\alpha}$-D-mannose 6-phosphate to ${\alpha}$-D-mannose 1-phosphate. The latter is the substrate for the synthesis of GDP-mannose, which serves as the mannosyl donor for many metabolic pathways in the cells. We report here on the isolation of a gene from a genomic library of Sphingomonas chungbukensis DJ77, the pmmC gene encoding phosphomannomutase. The gene was cloned into E. coli expression vector, and the sequence was analyzed. The ribosomal binding site GGAAG lays 5 bp upstream of the ORF of 750 bp, which is initiated by ATG codon and terminated by TAG. The predicted sequence of the enzyme consists of 249 amino acids with a molecular mass of 27.4 kDa and showed $86.9\%$ similarity to that of eukaryotic phosphomannomutase after bioinformatical analyses with the conserved domain search of NCBI. The purified gene product revealed the activity of phosphomannomutase. In conclusion, we confirmed that pmmC gene encodes phosphomannomutase actually.

Characteristics and Fabrication of Micro-Gas Sensors with Heater and Sensing Electrode on the Same Plane (동일면상에 heater와 감지전극을 형성한 마이크로가스센서의 제작 및 특성)

  • Lim, Jun-Woo;Lee, Sang-Mun;Kang, Bong-Hwi;Chung, Wan-Young;Lee, Duk-Dong
    • Journal of Sensor Science and Technology
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    • v.8 no.2
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    • pp.115-123
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    • 1999
  • A micro-gas sensor with heater and sensing electrode on the same plane was fabricated on phosphosilicate glass(PSG, 800nm)/$Si_3N_4$ (150nm) dielectric membrane. PSG film was provided by atmospheric pressure chemical vapor deposition(APCVD), and $Si_3N_4$ film by low pressure chemical vapor deposition (LPCVD). Total area of the fabricated device was $3.78{\times}3.78mm^2$. The area of diaphragm was $1.5{\times}1.5mm^2$, and that of the sensing layer was $0.24{\times}0.24mm^2$. Finite-element simulation was employed to estimate temperature distribution for a square-shaped diaphragm. The power consumption of Pt heater was about 85mW at $350^{\circ}C$. Tin thin films were deposited on the silicon substrate by thermal evaporation at room temperature and $232^{\circ}C$, and tin oxide films($SnO_2$) were prepared by thermal oxidation of the metallic tin films at $650^{\circ}C$ for 3 hours in oxygen ambient. The film analyses were carried out by SEM and XRD techniques. Effects of humidity and ambient temperature on the resistance of the sensing layer were found to be negligible. The fabricated micro-gas sensor exhibited high sensitivity to butane gas.

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