• 제목/요약/키워드: Noncoding DNA sequence

검색결과 34건 처리시간 0.032초

국내에서 분리된 canine parvovirus의 구조유전자 cloning과 염기서열 분석 (Cloning and sequencing of the gene encoding structural protein of canine parvovirus isolated in Korea)

  • 박종현;송재영;이중복;현방훈;안수환;전무형
    • 대한수의학회지
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    • 제32권4호
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    • pp.555-567
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    • 1992
  • In this study gene encoding structural proteins of a CPV isolate was cloned and sequenced to elucidate the molecular genetical properties of the canine parvoviruses isolated from the field. Six recombinant plasmids of pEP3, p1471, p2070, pEP069, pEP338 and p14711p were constructed from the map positions 22 to 98 of RF DNA to clone the VP1 and VP2 genes of CPV-V20. Sequentialy the gene comprising 3780 nucleotides were sequenced by dideoxy chain termination method. When nucleotide sequence of gene encoding the structural proteins of CPV-V20 was compared with those of other strains, CPV-N, CPV-d and CPV-780929 published previously, DNA, homologies to CPV-V20 were 99.87% with CPV-N, 99.73% with CPV-d, 96.85% with CPV-780929 and 98.4% with FPLV-Carl, respectively. The DNA sequence data of CPV-V20 showed seven point mutations and also deletion of 135 nucleotides from the nucleotide position 4745 to 4879 located in the 3'-noncoding region of CPV-N.

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최소자승 예측오차 확장 기반 가역성 DNA 워터마킹 (Least Square Prediction Error Expansion Based Reversible Watermarking for DNA Sequence)

  • 이석환;권성근;권기룡
    • 전자공학회논문지
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    • 제52권11호
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    • pp.66-78
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    • 2015
  • 바이오컴퓨팅 기술이 발전함에 따라 DNA 정보를 매개물로 한 DNA 워터마킹에 대한 연구가 이루어지고 있다. 그러나 DNA 정보는 일반 멀티미디어 데이터와는 달리 생물학적으로 중요한 정보이므로, 원본 DNA가 복원이 되는 가역성 DNA 워터마킹 기술이 필요하다. 본 논문에서는 최소자승 (Least square) 예측오차 확장 (prediction error expansion) 기반으로 비부호 DNA 서열의 가역성 워터마킹 기법을 제안한다. 제안한 방법에서는 비부호 영역의 4-문자 염기서열들을 인접한 개 염기에 의한 정수형 부호계수로 변환한다. 그리고 현재 부호계수에 대한 최소자승 예측 오차를 구한 다음, 예측오차 확장 조건에 따라 결정된 비트수만큼 예측오차를 확장한다. 이때 은닉된 인접 염기서열 간의 비교탐색을 통하여 허위개시코돈 생성을 방지한다. 실험 결과로부터 제안한 예측오차 확장 방법이 기존 방법과 평균 예측오차 확장 방법보다 높은 워터마크 용량을 가지며, 생물학적 변이 및 허위개시코돈이 발생되지 않음을 확인하였다.

Complete Chloroplast DNA Sequence from a Korean Endemic Genus, Megaleranthis saniculifolia, and Its Evolutionary Implications

  • Kim, Young-Kyu;Park, Chong-wook;Kim, Ki-Joong
    • Molecules and Cells
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    • 제27권3호
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    • pp.365-381
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    • 2009
  • The chloroplast DNA sequences of Megaleranthis saniculifolia, an endemic and monotypic endangered plant species, were completed in this study (GenBank FJ597983). The genome is 159,924 bp in length. It harbors a pair of IR regions consisting of 26,608 bp each. The lengths of the LSC and SSC regions are 88,326 bp and 18,382 bp, respectively. The structural organizations, gene and intron contents, gene orders, AT contents, codon usages, and transcription units of the Megaleranthis chloroplast genome are similar to those of typical land plant cp DNAs. However, the detailed features of Megaleranthis chloroplast genomes are substantially different from that of Ranunculus, which belongs to the same family, the Ranunculaceae. First, the Megaleranthis cp DNA was 4,797 bp longer than that of Ranunculus due to an expanded IR region into the SSC region and duplicated sequence elements in several spacer regions of the Megaleranthis cp genome. Second, the chloroplast genomes of Megaleranthis and Ranunculus evidence 5.6% sequence divergence in the coding regions, 8.9% sequence divergence in the intron regions, and 18.7% sequence divergence in the intergenic spacer regions, respectively. In both the coding and noncoding regions, average nucleotide substitution rates differed markedly, depending on the genome position. Our data strongly implicate the positional effects of the evolutionary modes of chloroplast genes. The genes evidencing higher levels of base substitutions also have higher incidences of indel mutations and low Ka/Ks ratios. A total of 54 simple sequence repeat loci were identified from the Megaleranthis cp genome. The existence of rich cp SSR loci in the Megaleranthis cp genome provides a rare opportunity to study the population genetic structures of this endangered species. Our phylogenetic trees based on the two independent markers, the nuclear ITS and chloroplast MatK sequences, strongly support the inclusion of the Megaleranthis to the Trollius. Therefore, our molecular trees support Ohwi's original treatment of Megaleranthis saniculifolia to Trollius chosenensis Ohwi.

소아의 Transfusion Transmitted Virus-Like Minivirus 유병률 (Prevalence of Transfusion Transmitted Virus-Like Mini Virus in Children)

  • 정주영;한태희
    • Pediatric Infection and Vaccine
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    • 제11권2호
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    • pp.153-157
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    • 2004
  • 목 적 : TTV는 인체 감염이 확인된 최초의 circovirus로 간염을 유발할 가능성에 대해 연구가 이루어지고 있다. TLMV는 최근에 발견된 circovirus로 TTV보다 작지만 유사한 구조를 가진 것으로 알려져 있다. TLMV 감염의 성인 유병률은 약 70%인 것으로 알려지지만 소아의 유병률은 아직 확실하지 않다. 이에 저자들은 국내 소아의 TLMV 유병률을 알아보기 위하여 시행하였다. 방 법 : 2001년 6월부터 12월까지 인제의대 상계 백병원 외래를 방문한 환아중 TTV DNA에 대한 PCR이 시행되었던 88명의 혈청 검체를 대상으로 하였다. TLMV의 5'NCR(noncoding region) 특이적 시발체를 이용하여 PCR을 시행하였다. 1라운드 PCR은 M1359, M1365 시발체를 사용하여 $94^{\circ}C$ 10분, $94^{\circ}C$에서 40초, $60^{\circ}C$에서 40초, $72^{\circ}C$에서 50초, $72^{\circ}C$에서 10분의 조건에서 55회 시행하였다. 최종 산물 $2{\mu}L$에 M1360, M1366 시발체를 사용하여 1라운드와 동일한 조건에서 PCR을 55회 시행하였다. TLMV PCR 양성이 나온 10건에 대해 직접 염기 서열 분석과 계통 분석을 시행하였다. 결 과 : 소아 전체 연령에서 TLMV 감염 유병률은 49%였다. 연령별 유병률은 생후 1세 미만은 36%, 1~3세는 62%, 4~6세는 43%, 7~9세는 16%, 10~15세는 66%였다. 전체 소아의 22%에서 TTV와 TLMV의 혼합 감염이 확인되었다. TLMV PCR 산물 10건에 대한 염기 서열 분석을 시행한 결과 다른 나라의 TLMV 염기 서열과 많은 차이가 났다. 결 론 : 국내 소아의 TLMV 유병률은 49%로 비교적 높았으며 TLMV와 TTV와 혼합 감염이 발생함을 알 수 있었다. 국내에서 유행하는 TLMV의 유전형이 다른 나라와 큰 차이가 있을 가능성이 있지만 이에 대한 연구가 더 필요할 것으로 보인다.

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A cDNA Clone for the 5' Exon of Chloroplast ATP Synthase Subunit I Gene (atpF) from Broccoli (Brassica oleracea L. var. Italica) and Its Expression Pattern

  • Choo Bong Hong
    • Journal of Plant Biology
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    • 제38권2호
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    • pp.137-141
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    • 1995
  • We isolated a cDNA clone, BLSC1, encoding 5' exon of ATP synthase CF0 subunit I from broccoli. BLSC1 is 285 nucleotides long which consists of a 5' noncoding region of 34 nucleotides, a 5' exon of 145 nucleotides and an intron of 106 nucleotides. The 5' exon codes for 48 amino acids which reveals mostly hydrophobic. The amino acid sequence deduced from BLSC1 shares 83%, 83% and 91% identities with the genes coding for atpF from wheat, rice and spinach, respectively. Genomic Southern blot analysis for BLSC1 showed a typically strong signal for a gene located in the chloroplast genome. Northern blot analysis identified three major classes of transcripts showing strong positive signals in the leaves, but only trace amounts of the transcripts were identified in the other organs like stems, flowr buds and roots.

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First report of Cycas necrotic stunt virus from cultivated Daphne plants

  • Lee, B.Y.;K.H. Ryu
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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    • pp.148.1-148
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    • 2003
  • Natural virus infection of cultivated Daphe odora plants showing chlorosis and stunting was observed and their causal agent was investigated. An isolate of isometic virus was purified from infected leaf tissues, and it could infect systemic severe mosaic on Chenopodium quinoa and C. amaranticolor. cDNA library was generated from partially purified viral RNAs and oligo dT primer-pSPORTl system, and recombinant clones were selected and their inserts were sequenced randomly. Nucleotide sequences of the virus were analyzed by BLAST, and it was closely related to members of subgroup B in the genus Nepovirus. The sequence analysis suggest that the virus was identified as an isolate of Cycas necrotic stunt virus (CNSV) because it was 89.7 % and 94.7 % identical to known CNSV for the CP and 3' noncoding region, respecitively. RT-PCR was performed to screen disease incidence of CNSV in Daphe plants, and five out of 10 plants (50 %) were infected by CNSV This is the first sequence information of CNSV from Daphe plants.

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Epigenetic regulation of long noncoding RNA UCA1 by SATB1 in breast cancer

  • Lee, Jong-Joo;Kim, Mikyoung;Kim, Hyoung-Pyo
    • BMB Reports
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    • 제49권10호
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    • pp.578-583
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    • 2016
  • Special AT-rich sequence binding protein 1 (SATB1) is a nuclear matrix-associated DNA-binding protein that functions as a chromatin organizer. SATB1 is highly expressed in aggressive breast cancer cells and promotes growth and metastasis by reprograming gene expression. Through genome-wide cross-examination of gene expression and histone methylation, we identified SATB1 target genes for which expression is associated with altered epigenetic marks. Among the identified genes, long noncoding RNA urothelial carcinoma-associated 1 (UCA1) was upregulated by SATB1 depletion. Upregulation of UCA1 coincided with increased H3K4 trimethylation (H3K4me3) levels and decreased H3K27 trimethylation (H3K27me3) levels. Our study showed that SATB1 binds to the upstream region of UCA1 in vivo, and that its promoter activity increases with SATB1 depletion. Furthermore, simultaneous depletion of SATB1 and UCA1 potentiated suppression of tumor growth and cell survival. Thus, SATB1 repressed the expression of oncogenic UCA1, suppressing growth and survival of breast cancer cells.

태백제비꽃군 ITS DNA 염기서열 분석 (Analysis of ITS DNA Sequences of the Viola albida Complex)

  • 황성수
    • 한국자원식물학회지
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    • 제19권5호
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    • pp.628-633
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    • 2006
  • 태백제비꽃군내 식물들은 동소적으로 생육하면서 단엽에서 장상복엽까지 연속적인 중간형을 나타내어 분류학적 어려움이 있다. 본 연구의 목적은 태백제비꽃, 단풍제비꽃, 남산제비꽃 그리고 각 분류군 사이의 중간형을 잎의 형태에 따라 5 집단으로 구분하고 각 집단별 대표적인 개체를 선별하여 ITS DNA 염기서열을 분석하고 분류학적 어려움을 해결하는데 있다. 정렬된 ITS1, ITS2 그리고 5.8S 지역의 염기서열은 702 bp로 나타났다. 5.8S 지역은 163 bp로 조사된 모든 개체에서 변이가 없었으며, ITS1과 ITS2는 일부 변이가 있어 분산분석, 염기 서열 분기 조사 그리고 분계분석에 이용하였다. 분산분석 결과 조사된 잎 형태별 개체들 간에 차이가 없는 것으로 나타났다. 염기서열 분기 조사 결과, 군외군으로 설정한 낚시제비꽃과 노랑제비꽃의 경우 Kimura 2-parameter distance에서 절대치가 0.05보다 훨씬 높게 나타나서 뚜렷한 차이가 확인되었다. 그러나 군내군 5 개체는 절대치가 모두 매우 낮게 나타나서 염기서열 분기는 종 수준이 아닌 종 이하의 수준으로 판단되었다. 분계분석에서 군외군으로 설정한 2 종은 기저 분계조를 형성하였다. 군내군은 하나의 분계조를 형성하였지만, 부트스트랩이 50% 이하로 나타나 계통학적 의미는 적은 것으로 사료된다.

Molecular Characterization and Infectious cDNA Clone of a Korean Isolate of Pepper mild mottle virus from Pepper

  • Yoon, Ju-Yeon;Hong, Jin-Sung;Kim, Min-Jea;Ha, Ju-Hee;Choi, Gug-Seon;Choi, Jang-Kyung;Ryu, Ki-Hyun
    • The Plant Pathology Journal
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    • 제21권4호
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    • pp.361-368
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    • 2005
  • A Korean isolate of Pepper mild mottle virus (PMMoV-Kr) was isolated from a diseased hot pepper plant and its biological and molecular properties were compared to that of PMMoV-J and PMMo V -So The genomic RNA of PMMoV-Kr consists of 6,356 nucleotides. The nucleotide and amino acid sequences identities of four viral proteins and two noncoding regions among PMMoV-Kr, PMMoV-S and PMMoV-J were $96.9\%\;to\;100.0\%\;and\;97.5\%\;to\;98.6\%$, respectively. Full-length cDNA amplicon of PMMoV-Kr was directly amplified by RT-PCR with a set of 5'-end primer anchoring T7 RNA promoter sequence and 3'-end virus-specific primer. Capped transcript RNAs from the full-length cDNA clone were highly infectious and caused characteristic symptoms of wild type PMMoV when mechanically inoculated to systemic host plants such as Nicotiana benthamiana and pepper plants.

Cloning and Expression of the Cathepsin F-like Cysteine Protease Gene in Escherichia coli and Its Characterization

  • Joo, Han-Seung;Koo, Kwang-Bon;Park, Kyun-In;Bae, Song-Hwan;Yun, Jong-Won;Chang, Chung-Soon;Choi, Jang-Won
    • Journal of Microbiology
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    • 제45권2호
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    • pp.158-167
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    • 2007
  • In this study, we have cloned a novel cDNA encoding for a papain-family cysteine protease from the Uni-ZAP XR cDNA library of the polychaete, Periserrula leucophryna. This gene was expressed in Escherichia coli using the T7 promoter system, and the protease was characterized after partial purification. First, the partial DNA fragment (498 bp) was amplified from the total RNA via RT-PCR using degenerated primers derived from the conserved region of cysteine protease. The full-length cDNA of cysteine protease (PLCP) was prepared via the screening of the Uni-ZAP XR cDNA library using the $^{32}P-labeled$ partial DNA fragment. As a result, the PLCP gene was determined to consist of a 2591 bp nucleotide sequence (CDS: 173-1024 bp) which encodes for a 283-amino acid polypeptide, which is itself composed of an 59-residue signal sequence, a 6-residue propeptide, a 218-residue mature protein, and a long 3'-noncoding region encompassing 1564 bp. The predicted molecular weights of the preproprotein and the mature protein were calculated as 31.8 kDa and 25 kDa, respectively. The results of sequence analysis and alignment revealed a significant degree of sequence similarity with other eukaryotic cysteine proteases, including the conserved catalytic triad of the $Cys^{90},\;His^{226},\;and\;Asn^{250}$ residues which characterize the C1 family of papain-like cysteine protease. The nucleotide and amino acid sequences of the novel gene were deposited into the GenBank database under the accession numbers, AY390282 and AAR27011, respectively. The results of Northern blot analysis revealed the 2.5 kb size of the transcript and ubiquitous expression throughout the entirety of the body, head, gut, and skin, which suggested that the PLCP may be grouped within the cathepsin F-like proteases. The region encoding for the mature form of the protease was then subcloned into the pT7-7 expression vector following PCR amplification using the designed primers, including the initiation and termination codons. The recombinant cysteine proteases were generated in a range of 6.3 % to 12.5 % of the total cell proteins in the E. coli BL21(DE3) strain for 8 transformants. The results of SDS-PAGE and Western blot analysis indicated that a cysteine protease of approximately 25 kDa (mature form) was generated. The optimal pH and temperature of the enzyme were determined to be approximately 9.5 and $35^{\circ}C$, respectively, thereby indicating that the cysteine protease is a member of the alkaline protease group. The evaluation of substrate specificity indicated that the purified protease was more active towards Arg-X or Lys-X and did not efficiently cleave the substrates with non-polar amino acids at the P1 site. The PLCP evidenced fibrinolytic activity on the plasminogen-free fibrin plate test.