• 제목/요약/키워드: ORF analysis

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Analysis of H3K4me3-ChIP-Seq and RNA-Seq data to understand the putative role of miRNAs and their target genes in breast cancer cell lines

  • Kotipalli, Aneesh;Banerjee, Ruma;Kasibhatla, Sunitha Manjari;Joshi, Rajendra
    • Genomics & Informatics
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    • 제19권2호
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    • pp.17.1-17.13
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    • 2021
  • Breast cancer is one of the leading causes of cancer in women all over the world and accounts for ~25% of newly observed cancers in women. Epigenetic modifications influence differential expression of genes through non-coding RNA and play a crucial role in cancer regulation. In the present study, epigenetic regulation of gene expression by in-silico analysis of histone modifications using chromatin immunoprecipitation sequencing (ChIP-Seq) has been carried out. Histone modification data of H3K4me3 from one normal-like and four breast cancer cell lines were used to predict miRNA expression at the promoter level. Predicted miRNA promoters (based on ChIP-Seq) were used as a probe to identify gene targets. Five triple-negative breast cancer (TNBC)-specific miRNAs (miR153-1, miR4767, miR4487, miR6720, and miR-LET7I) were identified and corresponding 13 gene targets were predicted. Eight miRNA promoter peaks were predicted to be differentially expressed in at least three breast cancer cell lines (miR4512, miR6791, miR330, miR3180-3, miR6080, miR5787, miR6733, and miR3613). A total of 44 gene targets were identified based on the 3'-untranslated regions of downregulated mRNA genes that contain putative binding targets to these eight miRNAs. These include 17 and 15 genes in luminal-A type and TNBC respectively, that have been reported to be associated with breast cancer regulation. Of the remaining 12 genes, seven (A4GALT, C2ORF74, HRCT1, ZC4H2, ZNF512, ZNF655, and ZNF608) show similar relative expression profiles in large patient samples and other breast cancer cell lines thereby giving insight into predicted role of H3K4me3 mediated gene regulation via the miRNA-mRNA axis.

자연산 대하(Fenneropenaeus chinensis)에서 검출된 노랑머리 바이러스 Genotype 8의 계통분류학적 특성 (Phylogenetic Characteristics of Yellow Head Virus (YHV) Genotype 8 Isolated from Fenneropenaeus chinensis in Korea)

  • 장광일;김보성;오윤경;황지연;권문경;김수미
    • 한국수산과학회지
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    • 제54권5호
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    • pp.698-702
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    • 2021
  • Yellow head virus (YHV) is a rod-shaped positive-sense single-stranded RNA virus, classified into the genus Okavirus, family Roniviridae, and order Nidovirales. In this study, 200 fleshy prawns (Fenneropenaeus chinensis) collected from the vicinity of Narodo in Goheung-gun, Korea, were screened for the presence of yellow head complex viruses and related genotype such as YHV genotype 8. The detection rate of YHV genotype 8 among the 200 fleshy prawns, determined using nested RT-PCR (reverse transcription polymerase chain reation), was 39.0%. Phylogenetic analysis of the ORF1b gene of YHV showed that eight distinct genetic lineages were detected. The four strains of YHV genotype 8 obtained in this study formed a robust clade with the YHV genotype 8 group that was first isolated from fleshy prawns in China suspected to have acute hepatopancreatic necrosis disease (AHPND).

한우 등심부위 근육 내 조지방함량에 따른 inducible cAMP early repressor (ICER) 유전자발현 분석 (Gene Expression Analysis of Inducible cAMP Early Repressor (ICER) Gene in Longissimus dorsi of High- and Low Marbled Hanwoo Steers)

  • 이승환;김남국;김성곤;조용민;윤두학;오성종;임석기;박응우
    • 생명과학회지
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    • 제18권8호
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    • pp.1090-1095
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    • 2008
  • 근내지방 축적이 왕성하게 진행되는 12개월 및 27개월령 사이에 유전자발현이 증가하는 inducible cAMP early repressor (ICER) 유전자의 클로닝 및 유전자발현 특성을 규명하기 위하여 한우조직별, 근육 내 조지방 함량에 따라 마블링 high- 및 low 한우 각 4두와 3두씩을 공시하였다. ddRT-PCR 분석을 통하여 증폭된 300 bp PCR 산물 및 EST 서열을 중심으로 1.5 kb 한우 ICER cDNA를 염기서열 분석하였으며, 조직별 유전자발현분석결과, 식이지방이 흡수되는 소장 조직에서 ICER 유전자의 발현량이 가장 높았다. 아울러, 같은 근육타입인 등심 및 우둔조직 사이에서 ICER 유전자발현은 등심조직에서 2.5배 많이 발현하였다. 한우 마블링 high, low 두 그룹 간 유전자발현 분석 결과, high 그룹에서 ICER 유전자발현이 4배 이상 증가하는 것으로 분석되었다. 따라서, ICER 유전자는 가축의 마블링과 밀접한 관련이 있는 유전자임이 사료된다.

Characterization of Xanthomonas axonopodis pv. glycines plasmids

  • Park, Sejung;Kim, Jung-Gun;Ingyu Hwang
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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    • pp.135.2-136
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    • 2003
  • To characterize plasmids in Xanthomonu axonopodis pv. glycines, we isolated plasmids pAG1 from the strain AG1 and pXAG81 and PXAG82 from the strain Bra, respectively, and sequenced three plasmids. The size of plasmids, pAG1, pXAG81, and pXAG82 was 15,149-base pairs (bp), 26,727-bp, and 1,496-bp, respectively Fifteen and twenty six possible open reading frames (ORFs) were present in pAG1 and pXAG81, respectively. Only one ORF homologous to a rep gene of Xylella fastidiosa was present in pXAG82. pAG1 contained genes homologous to avrBs3, tnpA, tnpR, repA, htrA, three parA genes, M.XmaI, R.XmaI, and six hypothetical proteins. pXAG81 contained genes homologous to avrBs3, tnpA, tnpR, repA, htrA, two parA genes, pemI, pemK, mobA, mobB, mobC, mobD, mobE, trwB, traF, traH, ISxac2, and eleven hypothetical proteins. Based on DNA sequence analysis, we presume that pXAG81 is a conjugal plasmid. Interestingly, we found 0.5-kb truncated avirulence gene similar to aurXacE3 on the right border of avrBs3 homolgs of pAG1 and pXAG81. Two hundred twenty five isolates were analyzed to find aurBS3 or tra gene homologs by Southern hybridization. The numbers of avrBs3 homolog varied from 3 in AG1 to 8 in AG166. Two hundred seventeen isolates appeared to can conjugative plasmids (pXAG81 type), and thirty eight isolates appeared to carry non-conjugative plamids (pAGl type). This indicated that aurBs3 gene homologs might be spread by conjugation in X. axonopodis pv. glycines.

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Expressed Sequence Tag Analysis for Identification and Characterization of Sex-Related Genes in the Giant Tiger Shrimp Penaeus monodon

  • Preechaphol, Rachanimuk;Leelatanawit, Rungnapa;Sittikankeaw, Kanchana;Klinbunga, Sirawut;Khamnamtong, Bavornlak;Puanglarp, Narongsak;Menasveta, Piamsak
    • BMB Reports
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    • 제40권4호
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    • pp.501-510
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    • 2007
  • Sex-related genes expressed in vitellogenic ovaries of the giant tiger shrimp, Penaeus monodon, were identified by an EST approach. A total of 1051 clones were unidirectionally sequenced from the 5 terminus. Nucleotide sequences of 743 EST (70.7%) significantly matched known genes previously deposited in the GenBank (E-value <$10^{-4}$) whereas 308 ESTs (29.3%) were regarded as newly unidentified transcripts (E-value >$10^{-4}$). A total of 559 transcripts (87 contigs and 472 singletons) were obtained. Thrombospondin (TSP) and peritrophin (79 and 87 clones accounting for 7.5 and 8.3% of clones sequenced, respectively) predominated among characterized transcripts. everal full length transcripts (e.g. cyclophilin, profillin and thioredoxin peroxidase) were also isolated. A gene homologue encoding chromobox protein (PMCBX, ORF of 567 nucleotides encoding a protein of 188 amino acids) which is recognized as a new member of the HP1 family was identified. Expression patterns of 14 of 25 sex-related gene homologues in ovaries and testes of P. monodon broodstock were examined by RT-PCR. Female sterile and ovarian lipoprotein receptor homologues were only expressed in ovaries whereas the remaining transcripts except disulfide isomerase related P5 precursor and adenine nucleotide translocator 2 were higher expressed in ovaries than testes of P. monodon broodstock. A homologue of ubiquitin specific proteinase 9, X chromosome (Usp9X) revealed a preferential expression level in ovaries than testes of broodstock-sized P. monodon (N = 13 and 11, P<0.05) but was only expressed in ovaries of 4-month-old shrimp (N = 5 for each sex).

Molecular Evidence of Recombination on Korean Isolates of Tomato yellow leaf curl virus by Nucleotide Transversions and Transitions

  • Lee, Hye-Jung;Park, Jung-An;Auh, Chung-Kyoon;Lee, Kyeong-Yeoll;Kim, Chang-Seok;Lee, Gwan-Seok;Soh, Hyun-Cheol;Choi, Hong-Soo;Lee, Suk-Chan
    • The Plant Pathology Journal
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    • 제27권4호
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    • pp.378-384
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    • 2011
  • Tomato yellow leaf curl virus (TYLCV), a member of genus Begomovirus, was isolated in Korea in 2008. We sequenced and analyzed the DNA-A of 51 TYLCV isolates from Korea, and 13 of the TYLCV isolates were selected as type representatives of TYLCV from six Korean provinces. The 13 TYLCV isolates were classified into Korea Group 1 (KG1, nine isolates) and Korea Group 2 (KG2, four isolates) based on the results of phylogenetic analysis and genome size (2774 and 2781 nucleotides, respectively). A recombination detection program 3 (RDP3) revealed two recombinations between the TYLCV Korea isolates and other TYLCV isolates [Thailand (AF206674), Iran (AJ132711), and Israel (X76319)]. TYLCV Jeju isolate was characterized by two recombination events (E1 and E2) caused by the presence of E1 in ORF V1 and C3, which may seem to be the mutations of the high nucleotide transversion and transition rate. Collectively, our results suggest that the occurrence of nucleotide transversions and transitions in TYLCV DNA-A might have induced novel recombination events within the TYLCV Korea isolates.

Characterization of dihydroflavonol 4-reductase cDNA in tea [Camellia sinensis (L.) O. Kuntze]

  • Singh, Kashmir;Kumar, Sanjay;Yadav, Sudesh Kumar;Ahuja, Paramvir Singh
    • Plant Biotechnology Reports
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    • 제3권1호
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    • pp.95-101
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    • 2009
  • Tea leaves are major source of catechins—antioxidant flavonoids. Dihydroflavonol 4-reductase (DFR, EC 1.1.1.219) is one of the important enzymes that catalyzes the reduction of dihydroflavonols to leucoanthocyanins, a key ''late'' step in the biosynthesis of catechins. This manuscript reports characterization of DFR from tea (CsDFR) that comprised 1,413 bp full-length cDNA with ORF of 1,044 bp (115-1,158) and encoding a protein of 347 amino acids. Sequence comparison of CsDFR with earlier reported DFR sequences in a database indicated conservation of 69-87% among amino acid residues. In silico analysis revealed CsDFR to be a membrane-localized protein with a domain (between 16 and 218 amino acids) resembling the NAD-dependent epimerase/dehydratase family. The theoretical molecular weight and isoelectric point of the deduced amino sequence of CsDFR were 38.67 kDa and 6.22, respectively. Upon expression of CsDFR in E. coli, recombinant protein was found to be functional and showed specific activity of 42.85 nmol $min^{-1}$ mg $protein^{-1}$. Expression of CsDFR was maximum in younger rather than older leaves. Expression was down-regulated in response to drought stress and abscisic acid, unaffected by gibberellic acid treatment, but up-regulated in response to wounding, with concomitant modulation of catechins content. This is the first report of functionality of recombinant CsDFR and its expression in tea.

Proteome Analysis of Paenibacillus polymyxa E681 Affected by Barley

  • Seul, Keyung-Jo;Park, Seung-Hwan;Ryu, Choong-Min;Lee, Yong-Hyun;Ghim, Sa-Youl
    • Journal of Microbiology and Biotechnology
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    • 제17권6호
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    • pp.934-944
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    • 2007
  • Paenibacillus polymyxa E681 is known to be able to suppress plant diseases by producing antimicrobial compounds and to promote plant growth by producing phytohormones, and secreting diverse degrading enzymes. In spite of these capabilities, little is known regarding the flow of information from the bacterial strain to the barley roots. In an attempt to determine the flow of information from the bacterial strain to barley roots, the strain was grown in the presence and absence of barley, and two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and MALDI-TOF mass spectrometry were used. 2D-PAGE detected approximately 1,000 spots in the cell and 1,100 spots in the supernatant at a pH 4-10 gradient. Interestingly, about 80 spots from each sample showed quantitative variations. Fifty-three spots from these were analyzed by MALDI-TOF mass spectrometry and 28 proteins were identified. Most of the cytosolic proteins expressed at higher levels were found in P. polymyxa E681 cells grown in the presence of barley rather than in the absence of barley. Proteins detected at a lower level in the surpernatant of P. polymyxa E68l cells grown in the presence of barley were lipoprotein, glucose-6-phosphate 1-dehydrogenase, heat-shock protein HtpG, spermidine synthase, OrfZ, ribonuclease PH, and coenzyme PQQ synthesis protein, and flagellar hook-associated protein 2 whereas proteins detected at a higher level in the surpernatant of P. polymyxa E681 cells grown in the presence of barley included D-alanyl-D-alanine ligase A, isopentenyl-diphosphate delta-isomerase, ABC transporter ATP-binding protein Uup, lipase. Many of the proteins belonging to plant-induced stimulons are associated with biosynthetic metabolism and metabolites of proteins and transport. Some of these proteins would be expected to be induced by environmental changes resulting from the accumulation of plant-secreted substances.

Cloning and Characterization of Cycloinulooligosaccharide Fructanotransferase (CFTase) from Bacillus polymyxa MGL21

  • Jeon, Sung-Jong;You, Dong-Ju;Kwon, Hyun-Ju;Shigenori Kanaya;Namio Kunihiro;Kim, Kwang-Hyeon;Kim, Young-Hee;Kim, Byung-Woo
    • Journal of Microbiology and Biotechnology
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    • 제12권6호
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    • pp.921-928
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    • 2002
  • Microorganism producing extracellular CFTase was isolated from soil and designated as Bacillus polymyxa MGL21. The gene encoding the CFTase (cft) from B. polymyxa MGL21 was cloned and sequenced. The ORF of the cf gene was composed of 3,999 nucleotides, encoding a protein (1,333 amino acids) with a predicted molecular mass of 149,375 Da. Sequence analysis indicated that CFTase was divided into five distinct regions. CFTase contained three regions of repeat sequences at the N-terminus and C-terminus. The endo-inulinase region of homology (ERH) of CFTase was similar to that of Pseudomonas mucidolens endo-inulinase ($50\%$ identity, 259 amino acids). Furthermore, CFTase possessed a highly conserved core region, which is considered to be functional for the hydrolysis reaction of inulin. The cft gene was expressed in a His-tagged form in Escherichia coli cells, and the His-tagged CFTase was purified to homogeneity. The optimal temperature and pH for CFTase activity were found to be $50^{\circ}C$ and 9.0, respectively. The enzyme activity was completely inhibited by 10 mM $Ag^+\;and\;Cu^2+$. Thin-layer chromatography analyses indicated that CFTase catalyzed not only the cyclization reaction ut also disproportionation and hydrolysis reactions as well.

조피볼락(Sebastes schlegelii) Interferon Regulatory Factor 8 (IRF8)의 분자유전학적 특성 및 발현 분석 (Molecular Characterization and Expression Analysis of Interferon Regulatory Factor 8 (IRF8) in the Black Rockfish Sebastes schlegelii)

  • 양혜림;권혁재;이성도;;김명진;이제희
    • 한국수산과학회지
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    • 제50권3호
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    • pp.302-310
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    • 2017
  • Interferon regulatory factor 8 (IRF8) is essential for the development of B and T cells, as well as for the activity of dendritic cells and macrophages. We performed molecular characterization of IRF8 from rock fish, Sebastes schlegelii (Ss), and investigated the spatial and temporal profile of mRNA expression after challenge with lipopolysaccharide (LPS), polyinosinic:polycytidylic acid (poly I:C), or Streptococcus iniae. The full-length cDNA sequence of SsIRF8 was 1,657 bp, containing an ORF of 1,266 bp. The gene had a predicted molecular mass of 47.7 kDa and an isoelectric point of 5.99. The amino acid sequence coded by this gene showed the highest degree of identity (90.8%) and similarity (96.2%) with IRF8 from Oplegnathus fasciatus. The SsIRF8 mRNA was expressed ubiquitously, at varying levels, with the highest level of expression observed in the spleen. To confirm the role of SsIRF8 in mediating the immune response, we measured SsIRF8 mRNA expression in the splenic tissue at different time points after injection with LPS, poly I:C, or S. iniae. The qRT-PCR results showed that SsIRF8 mRNA expression in the poly I:C-injected group was highly upregulated 6 hr after exposure (P<0.05). Expression of SsIRF8 mRNA in the S. iniae-injected group peaked at 24 hr. These results suggest that SsIRF8 might be important in regulating the strength of the rockfish immune response to immunostimulatory agents.