• Title/Summary/Keyword: promoter region sequencing

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A Deletion in Fungal Ras Promoter in Two Korean Strains of Oak Mushroom (Lentinula edodes)

  • Noh, Eun-Woon;Lee, Jae-Soon;Park, Young-Im;Park, Won-Chull
    • The Plant Pathology Journal
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    • v.18 no.2
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    • pp.74-76
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    • 2002
  • This study unexpectedly detected a deletion in the promoter region of ras gene in two Korean strains of oak mushrooms, Lentinula edodes (Berk.). Sequencing of the promoter regions revealed that one type consisting of two strains had a 113 bp deletion in the region. The pas promoter region of Korean strains differed by 16 bases from that of the Japanese strains. Between the two types of Korean strains, except for the deleted portion, only a single site appeared to be different.

A Study On the Application Methods of a Support Vector Machine for Gene Promoter Prediction. (유전자 프로모터 예측을 위한 Support Vector Machine의 응용 방법에 대한 연구)

  • Kim, Ki-Bong
    • Journal of Life Science
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    • v.17 no.5 s.85
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    • pp.714-718
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    • 2007
  • The high-throughput sequencing of a lot of genomes has resulted in the relatively rapid accumulation of an enormous amount of genomic sequence data. In this context, the problem posed by the detection of promoters in genomic DNA sequences via computational methods has attracted considerable attention in recent years since exact promoter prediction can give a clue to the elucidation of overall genetic networks. In this study, applications of support vector machine(SVM) to promoter prediction are explored to show a right approaches to discriminate between promoter and non-promoter regions by means of SVM. The results of various experiments show that encoding method, encoding region and learning data constitution can play an important role in the performance of SVM.

Computational Detection of Prokaryotic Core Promoters in Genomic Sequences

  • Kim Ki-Bong;Sim Jeong Seop
    • Journal of Microbiology
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    • v.43 no.5
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    • pp.411-416
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    • 2005
  • The high-throughput sequencing of microbial genomes has resulted in the relatively rapid accumulation of an enormous amount of genomic sequence data. In this context, the problem posed by the detection of promoters in genomic DNA sequences via computational methods has attracted considerable research attention in recent years. This paper addresses the development of a predictive model, known as the dependence decomposition weight matrix model (DDWMM), which was designed to detect the core promoter region, including the -10 region and the transcription start sites (TSSs), in prokaryotic genomic DNA sequences. This is an issue of some importance with regard to genome annotation efforts. Our predictive model captures the most significant dependencies between positions (allowing for non­adjacent as well as adjacent dependencies) via the maximal dependence decomposition (MDD) procedure, which iteratively decomposes data sets into subsets, based on the significant dependence between positions in the promoter region to be modeled. Such dependencies may be intimately related to biological and structural concerns, since promoter elements are present in a variety of combinations, which are separated by various distances. In this respect, the DDWMM may prove to be appropriate with regard to the detection of core promoter regions and TSSs in long microbial genomic contigs. In order to demonstrate the effectiveness of our predictive model, we applied 10-fold cross-validation experiments on the 607 experimentally-verified promoter sequences, which evidenced good performance in terms of sensitivity.

Subcloning and Sequencing of Maize rbcL Promoter Region

  • Woong-Seop Sim
    • Journal of Plant Biology
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    • v.38 no.1
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    • pp.107-113
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    • 1995
  • pRLYS1 containing intact rbcL gene of maize (Zea mays L. cv Golden X Bantam T-51; Zm-A) was digested with several restriction enzymes to construct subcones carrying promoter region of rbcL. The DNA fragments of 0.20, 0.19, 0.92 and 1.55 kb among the EcoRI digests, the EcoRI-DdeI digests, the AvaI digests and the EcoRI-BamHI digests of pRLYS1 were subcloned into pBluscriptSK+and named pRLPS2, pRLPS3, pRLPS14 and pRLPS35, respectively. Four subclones contain the 1.92 kb portion from 136 nucleotide downstream to 1780 nucleotide upstream from the ATG initiation codon of rbcL gene. pRLPS2 (-29 to -229) and pRLPS3 (-239 to -420 from the ATG) were sequenced. When nucleotide sequence of Zm-A was compared with sequence of rbcL promoter region of a different cultivar of maize (Zea mays L. cv WFG TMS X BS7; Zm-B), the difference rate between two cultivars was 4.3%. The mean of sequence divergence between Zm-A and three grass species in the same tribe, Andropogoneae, in the upstream region from 29 to 420 of ATG was 1.8%, whereas between Zm-B and above-mentioned three species was 5.4%. Therefore, Zm-A seems to evolutionarily closer to three other species in Andropogoneae tribe than Zm-B is.

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Genetic polymorphism in regulatory region of fatty acid binding protein 4 (FABP4) and its effect on carcass weight in Hanwoo steers (한우 지방산결합단백질 4(FABP4) 유전자 조절영역내 단일염기변이(SNP)와 도체형질간 연관성 분석)

  • Lee, Seung-Hwan;Kim, Nam-Kuk;Kim, Seung-Chang;Choi, Bong-Hwan;Heo, Kang-Neung;Lee, Chang-Soo;Kim, Oun-Hyun;Lee, Jun-Heon;Kim, Hyeong-Cheul;Hong, Seong-Koo
    • Korean Journal of Agricultural Science
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    • v.38 no.4
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    • pp.673-680
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    • 2011
  • The aim of this study was to identify the polymorphism on fatty acid binding protein (FABP4) gene promoter region and its association with carcass traits in Hanwoo. We performed PCR-direct sequencing of FABP4 promoter region to identify single nucleotide polymorphism (SNPs) using unrelated 24 Hanwoo bulls. Four SNPs (-298A>G, -472A>G, -887A>G, -862A>G) were detected in the promoter region and genotyped on 583 Hanwoo steers. A linear mixed model revealed an association of three SNPs (-298A>G, -472A>G and -862A>G) with carcass weight and marbling score in dominance model (P<0.05). The animals with AA genotypes for the three SNPs were heavier carcass weight (5 kg) than animals with GG genotypes in the statistical analysis. For the marbling score, the AA genotype was lower effect of marbling score (0.21) than GG genotypes. In conclusion, this study indicates an important role for three SNPs detected in promoter region of FABP4 in determining carcass weight and marbling score in Hanwoo.

사람의 Serine palmitoryl transferase II 및 ceramidase의 promoter에 대한 연구

  • Kim, Hui-Suk;Song, Seong-Gwang;Lee, Eun-Yeol;Lee, Sang-Do;Linn, Steve;Merrill, Alfred H.
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.588-591
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    • 2000
  • Serine palmitoyl transferase(SPT) and ceramidase are the key enzymes in sphingolipid biosynthesis. To study sphingolipid metabolism, we have got the 5'-upstream regions of human serine palmitoyl transferase subunit II and acid ceramidase gene by using GenomeWalker kits(Clontech Co.). Human genomic DNA was purified from HT29, human colon canser cell line by using DNAzol. We got several bands after secondary PCR and subcloned them to T7bule vector. Human SPTII promoter which we got was 2690bp but we cut it with Bgl II and vector with Bgl II and BamH I, and subcloned 1782bp to pGL2-enhancer vector and pGL2-basic vector with luciferase reporter gene. Human acid ceramidase promoter which we got were 2028bp and 1034bp and subcloned to pGL2-enhancer vector and pGL2-basic vector. We transfected these promoters to HT29 cell and assayed luciferase activity. For measuring transfection efficiency, pRL-TK vector with seapancy luciferase reproter gene was cotransfected with these promoters.

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Analysis of Tandem Repeats in the Promoter Region of iNOS Gene in Korean Genome

  • Kim, Sun-Ji;Yoo, Min
    • Biomedical Science Letters
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    • v.14 no.2
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    • pp.127-130
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    • 2008
  • To investigate if there are tandem repeats in iNOS gene in Korean genome we applied PCR amplification followed by DNA sequencing. Tandem repeats we were looking at were (AAAT)n in the promoter region. Totally, 65 people were subjected for this experiment. Twenty of them were patients with metabolic disease. Only $(AAAT)_4$ was found in all of these Korean samples. This result was somewhat different trom the data for Caucasians and other Asian people. So, we assume this is specific VNTR (variable number of tandem repeat) in Korean which can be used for the purpose of diagnosis and for the differentiation of ethnic groups.

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Effect of random Shine-Dalarno sequence on the expression of Bovine Growth Hormone Gene in Escherichia coli (대장균에서 무작위 샤인-달가노 서열이 소성장호르몬 유전자 발현에 미치는 영향)

  • 나경수;나경수;백형석;이용세
    • Journal of Life Science
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    • v.10 no.4
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    • pp.422-430
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    • 2000
  • In order to search for the effects of Shine-Dalgarno (SD) sequence and nucleotide sequence of spacer region (SD-ATG) on bGH expression, oligonucleotides containing random SD sequences and a spacer region were chemically synthesized. The distance between SD region and initiation codon (ATG) was fixed to 9 nucleotides in length. The expression vectors have been constructed using pT7-1 vector containing a T7 promoter. Positive clones were screened with colony hybridization and named pT7A or pT7B plasmid series. The selected clones were confirmed by DNA sequencing and finally, 19 clones having various SD combinations were obtained. When bovine growth hormone was induced by IPTG in E. coli BL21(DE3), all cells harboring these plasmids produced a detectable level of bGH in western blot analysis. However, various SD sequences did not affect on bGH expression, indicating that the sequences of SD and the spacer region did not sufficiently destabilize mRNA secondary structure of bGH gene. Therefore, these results indicate that the disruption of mRNA secondary structure might be a major factor for regulating bGH expression in the translational initiation process.

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Analysis of Upstream Regulatory Region from Populus nigra × P. maximowiczii by Inverse PCR Technique (Inverse PCR 기법(技法)을 이용(利用)한 양황철 DNA의 Regulatory Region의 탐색(探索))

  • Son, Suk Gyu;Hyun, Jung Oh
    • Journal of Korean Society of Forest Science
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    • v.87 no.3
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    • pp.334-340
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    • 1998
  • This research was conducted to identify plant regulatory regions by gene tagging method. A promoterless GUS coding sequence was introduced to Populus nigra ${\times}$ P. maximowiczii via Agrobacterium strains(LBA4404/EHA101), and putative transgenic poplars were selected by culturing on medium containing G418($60mg/{\ell}$) and by GUS assay. Among them one positive plant was to amplify the native sequences flanking to the introduced GUS gene in plant genome by inverse PCR method and from this 730 by DNA product was obtained. After subcloning and sequencing, it has 88% homology to the Eucalyptus gunnii CAD(cinnamyl alcohol dehydrogenase) gene. The GUS gene fused with the putative promoter reinserted into poplar leaves by particle bombardment method to test the funtional promoter activity. Upon staining with X-gluc, many blue spots appeared on the leaf segments bombarded by the chimeric gene 2-3 days, thus the isolated DNA fragment contain some possible coding region as well as a putative regulatory sequences of poplar CAD gene.

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