• Title/Summary/Keyword: URA3

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Characterization of a New Gene Resistant to Alkylating Agents and 3-Aminobenzamide When Knocked Out in Fission Yeast (분열형 효모에서 유전자 결실에 의해 알킬화제와 3-AMINOBENZAMIDE에 저항성을 나타내는 새로운 유전자의 특성 분석)

  • 박종군;차재영;황성진;박세근;김미영;백성민;최인순;이정섭
    • Journal of Life Science
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    • v.12 no.2
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    • pp.219-225
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    • 2002
  • The organization of eukayotic chromatin into specific conformation that are associated with transcription, replication, reapir and other nuclear processes are achieved via a series of DNA-protein interaction. These interactions are mediated by a range of DNA-binding domains such as SAP domain et at. By searching S. pombe genomic DNA database, we have found a gene named SAPuvs (SAP UV Sensitive) whose amino acid sequence is in part similar to SAP domain of Arabidopsis poly (ADP-ribose) polymerase and Ku7O. Knock-out cell of S. pombe SAPuvs gene was constructed using Ura4 as a selection marker. Survival analysis of knock-out cell indicated that treatment with UV significantly reduces the survival compared to wild type cell. Potentiation of MMS-induced cytotoxicity by 3AB post-treatment was observed in wild type cells, but not in knock-out cells. These data suggested that the protein encoded by SAPuvs gene is associated with chromatin reorganization during DNA repair.

Characterization of the cloned RNA1 gene of Saccharomyces cerevisiae (Cloning된 효모의 RNAI 유전자의 특성에 관하여)

  • Song, Young-Hwan;Kim, Dae-Young;Kim, Jin-Kyung
    • Journal of fish pathology
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    • v.6 no.2
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    • pp.93-101
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    • 1993
  • The RNAI mutation of Saccharomyces cerevisia is a recessive and temperature sensitive lethal mutation which interferes with the production of mRNA, rRNA, and tRNA. However, the precise role of RNAI gene have not been revealed until yet. We have cloned rna1-1 mutant gene from rna1-1 mutant yeast strain(R49 ; trpl, ura3-52, rna1-1). The 3.4kb BglII fragment of wild type RNAI clone(81-2-6) contains whole RNAI gene. The genomic southern blotting with BglII digested R49 genomic DNA as a probe shows the unique and identical band with wild type 3.4kb BglII fragment. Therefore, We prepared partial BglII genomic library(3~4kb BglII fragments) into BamH I site of pUC19. The rna 1-1 mutant clone was screened with Digoxigenin(DIG)-lableled probe by high density colony hybridization. The 5'-flanking region of rna1-1 gene was sequenced by dideoxy chain termination method. The 5'-flanking sequence of RNAI gene contains three TATA-like sequence ; TAATA, TATA and TTTTAA at position of -67, -45, and -36 from first ATG codon respectively. The 5'-flanking region of wild type RNA I gene from ATG codon to -103nt was deleted with Bal31 exonuclease digestion, generating $pUC{\Delta}$/RNA I. After constructing $pYEP{\Delta}RNA$ I (consists of -103nt deleting RNA I gene, URA3 gene, $2{\mu}m$ rep. origin), pYEPrna1-1(consists of Xba I fragment of pUCrna1-1. URA3 gene, $2{\mu}m$ rep. origin), and pYEPRNAI. each plasmid was transformed into host strain(trpl, ura3-52, rna1-1) by electroporation, respectively. Yeast transformant carrying $pYEP{\Delta}RNA$ I did not complement the thermal sensitivity of rna1-1 gene. It means that TATA-like sequences in 5'-flanking region is not TATA sequence for transcribing RNAI gene and there may be other essential sequence in upstream region for the transcription of RNAI gene.

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Expression of Recombinant Korean Mistletoe(KM) Lectin and B genes in Saccharomyces cerevisiae (Saccharomyces cerevisae에서 한국산 겨우살이 유래 lectin A 및 B 유전자의 발현)

  • 최윤혁;김종배;양웅석;황철원
    • Journal of Life Science
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    • v.14 no.5
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    • pp.840-846
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    • 2004
  • A study for expression of Korean Mistletoe (KM) lectin gene (A,B) in Saccharomyces cerevisiae was done using transforming system of yeast. In order to overexpress the genes efficiently in yeast, two lectin genes (A,B) were re-cloned and modified including Kozak translation initiation sequence using PCR amplification. The constructed plasmids containing modified lectin A and B genes were transformed to S. cerevisea INVSc (MAT G, his3 $\Delta$1, leu2, trpl-289, ura3-52). The transformed cells were identified by DNA sequencing with ABI3700 system and induced with 2% of galactose for recombinant KM lectin (rKM lectin) protein. The rKM lectin A and B proteins were determinated about 29kDa size of protein by SOS-P AGE and western blotting analysis. The expressed recombinant lectin was determinated 1.24∼1.75 $\mu\textrm{g}$ per 1 mg of cytosolic soluble protein by sandwich ELISA method. Moreover the lectin genes were expressed as maximum level at 36 h after galactose induction and lectin A gene was were repressed after 48 h.

Classification of Needle Type and Growth Characteristics of Cryptomeria japonica Planted in Cheju Province (삼나무의 엽형분류(葉型分類) 및 생장특성(生長特性)에 관(關)한 연구(硏究) -제주도(濟州道)를 중심(中心)으로-)

  • Jin, Hyun O;Chon, Sang-Keun
    • Journal of Korean Society of Forest Science
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    • v.76 no.4
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    • pp.410-417
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    • 1987
  • Needle type and growth characteristics of Cryptomeria japonica planted in Cheju, Korea, were investigated and compared with those of the same species distributed in Japan. Needle types were classified into nine lines, and the stands were composed of more than six lines. The trees of the same needle type as Ura-Sugi (Cryptomeria japonica) in Japan showed the best growth in height.

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Interpolation Technique for 3-D Conformal Array (3차원 콘포멀 어레이에서의 인터폴레이션 기술의 적용)

  • Kang, Kyung-mook;Seol, Kyung-Eun;Jeon, Junghwan;Koh, Jinhwan
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.41 no.12
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    • pp.1748-1751
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    • 2016
  • In this correspondence, we studied 3D uniform rectangular array as an extension of interpolation technique to compensate the beam pattern of 3D conformal array. The simulation result shows outstanding performance comparing to 2D interpolations.

3-D interpolation technique and compressive sensing for 3-D conformal array (3차원 interpolation technoque과 compressive sensing을 이용한 비 균일한 3차원 array의 beam pattern 복구)

  • Kang, K;Seol, K;Cesar, W;Jeong, S;Koh, J
    • Proceedings of the Korea Information Processing Society Conference
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    • 2017.04a
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    • pp.106-108
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    • 2017
  • 본 논문에서는 휘어지거나 굴곡진 array인 3차원 conformal array의 beam pattern을 보정하고자 기존의 2차원에서 3차원으로 확장한 interpolation technique과 compressive sensing을 이용하여 3-D uniform rectangular array(3-D URA)에 적용하는 방법을 연구하였다. 시뮬레이션 결과는 compressive sensing이 interpolation technique보다 우수한 특성을 보여준다.

A Pyrenyl-Appended Triazole-Based Calix[4]arene as a Fluorescent Sensor for Iodide Ion

  • Kim, Jong-Seung;Park, Sun-Young;Kim, Sang-Hoon;Thuery, Pierre;Souane, Rachid;Matthews, Susan E.;Vicens, Jacques
    • Bulletin of the Korean Chemical Society
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    • v.31 no.3
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    • pp.624-629
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    • 2010
  • The synthesis and evaluation of a novel calix[4]arene-based fluorescent chemosensor 1 for the detection of I. is described. The fluorescent changes observed upon addition of various anions show that 1 is selective for I. over other anions. Addition of I. results in ratiometric measurements with 1 : 1 complex ratio.

In vitro Biodegradability and Surface Properties of Block Copoly(ester-ether)s Consisting of Poly(L-lactide) and Polyether

  • Lee, Chan-Woo;Kim, Yoshiharu ura
    • Macromolecular Research
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    • v.11 no.1
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    • pp.42-46
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    • 2003
  • Cell attachment and proliferation on the polymer films of triblock copolymer(ester-ether)s comprising po1y (L-1actide) (PLLA) and poly (oxyethylene-co-oxypropylene)(PN) were investigated using 3T3 fibroblasts. It was found that on the tissue culture polystyrene(TCPS) and the PLLA control film the cells could spread well while on the copolymer films the cells showed a rounded morphology without spreading and proliferated weakly. Especially, little cells proliferated on the films of copolymer having a LN composition of 20 wt%. While the water absorption of the copolymer films increased with increasing PN content, the contact angle against water of copolymer films immersed in aqueous medium was almost identical, being slightly lower than that of the PLLA film. These properties were compatible with the results of cell attachment. The in vitro hydrolysis of the films of triblock and multiblock type copolymers was faster with increasing PN content. The increased hydrolyzability, the flexibility and the decreased cell attachment suggested that these copolymers may have high potential as biodegradable materials for medical use.

Cloning and Characterization of Replication Origins from Misgurnus mizolepis (미꾸라지로부터의 복제원점 클로닝 및 그 특성에 관한 연구)

  • Lim Hak-Seob;Kim Moo-Sang;Lee Hyung-Ho
    • Journal of Aquaculture
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    • v.8 no.3
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    • pp.209-220
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    • 1995
  • The nuclear matrix was isolated from Misgumus mizolepis liver nuclei by low salt extraction and restriction enzyme treatment. The structure was digested with proteinase K. After centrifugation, matrix attachment regions (MARs) were obtained by RNase treatment and phenol-chloroform extraction. The result leads to the appearance of smeared bands in the range of about 0.3-15 kb. pURY19 vector was constructed by inserting 2.13 kb Eco47 III fragment of the yeast uracil 3 gene into the unique Ssp I site of pUC19 plasmid vector as a selection marker. This vector is unable to be maintained in Sacrharomyces cerevisiae by itself since it cannot replicate as an extrachromosomal element. Using this system, we attempted cloning the ARS (autonomously replicating sequence) from M. mizelepis to develop an efficient expression vector for the transgenic fish. pURY19N_{l-62}$ were constructed by inserting MARs in pURY19 plasmid vector and transformation of E. coli $DH5\alpha$. Replication origins (ARS) of M. mizolepis were isolated, which enabled the vector to replicate autonomously in S. cerevisiae. The cloned DNA fragments were sequenced by Sanger's dideoxy-chain termination method. All clones were AT-rich. $pURY19N_6$, one of the clones, expecially contained ARS consensus sequence, Topoisomerase II consensus, near A-box and T-box.

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Genetic Variations of Candida glabrata Clinical Isolates from Korea using Multi-locus Sequence Typing (Multi-locus sequence typing을 이용한 한국에서 분리한 Candida glabrata 임상균주의 유전자 유형 분석)

  • Kang, Min Ji;Lee, Kyung Eun;Jin, Hyunwoo
    • Journal of Life Science
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    • v.30 no.2
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    • pp.122-128
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    • 2020
  • Although Candida albicans is the major fungal pathogen of candidemia, severe infections by non-albicans Candida (NAC) spp. have been increasing in recent years. Among NAC spp., C. glabrata has emerged as the second most common pathogen. However, few studies have been conducted to investigate its structure, epidemiology, and basic biology. In the present study, multi-locus sequence typing (MLST) was performed with a total of 102 C. glabrata clinical isolates that were isolated from various types of clinical specimen. For MLST, six housekeeping genes-FKS, LEU2, NMT1, TRP1, UGP1, and URA3-were amplified and sequenced. The results were analyzed using the C. glabrata database. Out of a total of 3,345 base-pair DNA sequences, 49 variable nucleotide sites were found, and the results showed that 12 different sequence types (ST) were identified from the 102 clinical isolates. The data also demonstrated that the undetermined ST1 was the most predominant ST in Korea. Further, seven undetermined STs (UST) containing UST2-8 were classified at specific loci. The data from this study may provide a fundamental database for further studies on C. glabrata, including its epidemiology and evolution. The data may also contribute to the development of novel antifungal agents and diagnostic tests.