• Title/Summary/Keyword: conserved sequences

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Marker Production by PCR Amplification with Primer Pairs from Conserved Sequences of WRKY Genes in Chili Pepper

  • Kim, Hyoun-Joung;Lee, Heung-Ryul;Han, Jung-Heon;Yeom, Seon-In;Harn, Chee-Hark;Kim, Byung-Dong
    • Molecules and Cells
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    • v.25 no.2
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    • pp.196-204
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    • 2008
  • Despite increasing awareness of the importance of WRKY genes in plant defense signaling, the locations of these genes in the Capsicum genome have not been established. To develop WRKY-based markers, primer sequences were deduced from the conserved sequences of the DNA binding motif within the WRKY domains of tomato and pepper genes. These primers were derived from upstream and downstream parts of the conserved sequences of the three WRKY groups. Six primer combinations of each WRKY group were tested for polymorphisms between the mapping parents, C. annuum 'CM334' and C. annuum 'Chilsung-cho'. DNA fragments amplified by primer pairs deduced from WRKY Group II genes revealed high levels of polymorphism. Using 32 primer pairs to amplify upstream and downstream parts of the WRKY domain of WRKY group II genes, 60 polymorphic bands were detected. Polymorphisms were not detected with primer pairs from downstream parts of WRKY group II genes. Half of these primers were subjected to $F_2$ genotyping to construct a linkage map. Thirty of 41 markers were located evenly spaced on 20 of the 28 linkage groups, without clustering. This linkage map also consisted of 199 AFLP and 26 SSR markers. This WRKY-based marker system is a rapid and simple method for generating sequence-specific markers for plant gene families.

MOTIF BASED PROTEIN FUNCTION ANALYSIS USING DATA MINING

  • Lee, Bum-Ju;Lee, Heon-Gyu;Ryu, Keun-Ho
    • Proceedings of the KSRS Conference
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    • v.2
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    • pp.812-815
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    • 2006
  • Proteins are essential agents for controlling, effecting and modulating cellular functions, and proteins with similar sequences have diverged from a common ancestral gene, and have similar structures and functions. Function prediction of unknown proteins remains one of the most challenging problems in bioinformatics. Recently, various computational approaches have been developed for identification of short sequences that are conserved within a family of closely related protein sequence. Protein function is often correlated with highly conserved motifs. Motif is the smallest unit of protein structure and function, and intends to make core part among protein structural and functional components. Therefore, prediction methods using data mining or machine learning have been developed. In this paper, we describe an approach for protein function prediction of motif-based models using data mining. Our work consists of three phrases. We make training and test data set and construct classifier using a training set. Also, through experiments, we evaluate our classifier with other classifiers in point of the accuracy of resulting classification.

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Phylogenetic Analysis of Caterpillar Fungi by Comparing ITS 1-5.8S-ITS 2 Ribosomal DNA Sequences

  • Park, Joung-Eon;Kim, Gi-Young;Park, Hyung-Sik;Nam, Byung-Hyouk;An, Won-Gun;Cha, Jae-Ho;Lee, Tae-Ho;Lee, Jae-Dong
    • Mycobiology
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    • v.29 no.3
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    • pp.121-131
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    • 2001
  • This study was carried out to identify the phylogenetic relationships among several caterpillar fungi by comparing the sequences of internal transcribed spacer regions(ITS1 and ITS2) and 5.8S ribosomal DNA(rDNA) repeat unit. The sequences of ITS1, ITS2, and the 5.8S rDNA from 10 strains of Cordyceps species, 12 strains of Paecilomyces, 3 strains of Beauveria, 2 strains of Metarhizium and 1 strains of Hirsutella were amplified, determined and compared with the previously known Cordyceps species. The sequences of 5.8S rDNA were more conserved in length and variation than those of ITS regions. Although the variable ITS sequences were often ambiguously aligned, the conserved sites could be found. In the phylogenetic tree, the species generally divided into three clusters, supported by their morphology and/or host ranges. The 5.8S rDNA and TTS1 sequences among 10 species of Cordyceps militaris were identical and only one base pair in ITS2 sequence was different. Cordyceps sinensis and Cordyceps ophioglossoides were also clearly different, although they belonged to the same cluster. The Geniank database search of species revealed sister taxa of an entomogenous fungus. Metarhizium was used as an putgroup in all taxa.

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A Subpopulation of RNA3 of Cucumber mosaic virus Quasispecies

  • Park, Seung-Kook;Park, Sun-Hee;Yoon, Ju-Yeon;Park, Jang-Kyung;Ryu, Ki-Hyun
    • The Plant Pathology Journal
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    • v.19 no.4
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    • pp.210-216
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    • 2003
  • This study examined the existence of genetically diverse population of Cucumber mosaic virus (CMV), known as quasispecies, from lily, Nicotiana benthamiana and from purified virions. Based on the conserved sequences of CMV lily isolates in intergenic region (IR) on RNA3, the genetic variation of IR from three different sources was investigated by a specific restriction endonuclease hydrolysis of amplified reverse transcription-polymerase chain reaction (RT-PCR) products using virus-specific primers, and was compared with IR sequences. The IR nucleotide sequences of CMV lily isolates were highly conserved, however, quasispecies was detected from all three sources in low level, containing sub-populations of RNA3. These subpopulations of RNA3 were inoculated onto zucchini squash by in vitro transcripts from corresponding full-length cDNA clones together with Eny RNA1 and 2 transcripts. The systemic symptom of zucchini plants infected by these quasispecies was chlorotic spotting, which was milder than severe mosaic and stunt symptom caused by Eny-CMV. The severity of symptom was correlated with RNA accumulation of viruses. These results suggest that the genome of CMV lily isolates consists of quasispecies populations.

Regulatory Sequences in the 5' Flanking Region of Goat β-Casein Gene

  • Huang, Mu-Chiou;Chao, Jiunn-Shiuan
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.11
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    • pp.1628-1633
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    • 2001
  • A goat ${\beta}$-casein gene was cloned and sequenced. Our previous study had determined the nucleotide sequences of the 5' flanking region and the structural gene including all 9 exons. In the present study, investigations were done on the regulatory sequences in the 5' flanking region of the goat ${\beta}$-casein gene by aligning and comparing it with the same gene from other mammals. The results showed that -200/-1 bp of the 5' flanking sequences contained six conserved clusters, in which the sites of gene expression regulated by the transcription factor and hormone might exist. It showed that fourteen glucocorticoid receptor elements, two cAMP responsive elements, two SV40 virus enhancer core sequences, two OCT-1 binding elements and one CTF/NF-1 binding element were dispersed in the 5' flanking region of goat ${\beta}$-casein gene. Our findings are perhaps valuable for the elucidation of the molecular mechanisms that control the expression of the goat ${\beta}$-casein gene.

Odorant Receptors Containing Conserved Amino Acid Sequences in Transmembrane Domain 7 Display Distinct Expression Patterns in Mammalian Tissues

  • Ryu, Sang Eun;Shim, Tammy;Yi, Ju-Yeon;Kim, So Yeun;Park, Sun Hwa;Kim, Sung Won;Ronnett, Gabriele V.;Moon, Cheil
    • Molecules and Cells
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    • v.40 no.12
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    • pp.954-965
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    • 2017
  • Mammalian genomes are well established, and highly conserved regions within odorant receptors that are unique from other G-protein coupled receptors have been identified. Numerous functional studies have focused on specific conserved amino acids motifs; however, not all conserved motifs have been sufficiently characterized. Here, we identified a highly conserved 18 amino acid sequence motif within transmembrane domain seven (CAS-TM7) which was identified by aligning odorant receptor sequences. Next, we investigated the expression pattern and distribution of this conserved amino acid motif among a broad range of odorant receptors. To examine the localization of odorant receptor proteins, we used a sequence-specific peptide antibody against CAS-TM7 which is specific to odorant receptors across species. The specificity of this peptide antibody in recognizing odorant receptors has been confirmed in a heterologous in vitro system and a rat-based in vivo system. The CAS-TM7 odorant receptors localized with distinct patterns at each region of the olfactory epithelium; septum, endoturbinate and ectoturbinate. To our great interests, we found that the CAS-TM7 odorant receptors are primarily localized to the dorsal region of the olfactory bulb, coinciding with olfactory epithelium-based patterns. Also, these odorant receptors were ectopically expressed in the various non-olfactory tissues in an evolutionary constrained manner between human and rats. This study has characterized the expression patterns of odorant receptors containing particular amino acid motif in transmembrane domain 7, and which led to an intriguing possibility that the conserved motif of odorant receptors can play critical roles in other physiological functions as well as olfaction.

Identification of Novel Cupredoxin Homologs Using Overlapped Conserved Residues Based Approach

  • Goyal, Amit;Madan, Bharat;Hwang, Kyu-Suk;Lee, Sun-Gu
    • Journal of Microbiology and Biotechnology
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    • v.25 no.1
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    • pp.127-136
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    • 2015
  • Cupredoxin-like proteins are mainly copper-binding proteins that conserve a typical rigid Greek-key arrangement consisting of an eight-stranded β-sandwich, even though they share as little as 10-15% sequence similarity. The electron transport function of the Cupredoxins is critical for respiration and photosynthesis, and the proteins have therapeutic potential. Despite their crucial biological functions, the identification of the distant Cupredoxin homologs has been a difficult task due to their low sequence identity. In this study, the overlapped conserved residue (OCR) fingerprint for the Cupredoxin superfamily, which consists of conserved residues in three aspects (i.e., the sequence, structure, and intramolecular interaction), was used to detect the novel Cupredoxin homologs in the NCBI non-redundant protein sequence database. The OCR fingerprint could identify 54 potential Cupredoxin sequences, which were validated by scanning them against the conserved Cupredoxin motif near the Cu-binding site. This study also attempted to model the 3D structures and to predict the functions of the identified potential Cupredoxins. This study suggests that the OCR-based approach can be used efficiently to detect novel homologous proteins with low sequence identity, such as Cupredoxins.

Investigation of Conserved Regions in Lipase Genes (Lipase 유전자의 보존적 영역 탐색)

  • 이동근;김철민;김상진;이상현;이재화
    • Journal of Life Science
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    • v.13 no.5
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    • pp.723-731
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    • 2003
  • For the investigation of conserved regions in lipase genes, 132 and 24 sequences were obtained from LED (Lipase Engineering Database) and COG (Clusters of Orthologous Groups of proteins), respectively. There was high diversity in lipase genes and peculiar amino acid sequences were conserved for each homologous family of LED. Similar conserved amino acid sequences were detected from COG0657 and Moraxella lipase 1 homologous group of LED. Although many studies have attempted to detect new lipase genes in procaryotes, they have been limited culturable bacteria. The importance of metagenome, including DNA from non-culturable bacteria, is known. Due to the high diversity, we assumed it might be possible to detect new lipase gene from metagenome. Due to the high diversity of nucleotide sequences in lipase genes, 10 primer sets were designed. Designed primer sets were inspected in BLAST of NCBI and they could amplify a part of the lipase gene from 222 to 713 bp. They can amplify 16.7%∼60.0% of each lipase homologous group which was 3.6 fold higher than each sets. They might offer a high probability of detecting new lipase genes, owing to high efficiency and the diversity of lipase genes.

Insilico profiling of microRNAs in Korean ginseng (Panax ginseng Meyer)

  • Mathiyalagan, Ramya;Subramaniyam, Sathiyamoorthy;Natarajan, Sathishkumar;Kim, Yeon Ju;Sun, Myung Suk;Kim, Se Young;Kim, Yu-Jin;Yang, Deok Chun
    • Journal of Ginseng Research
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    • v.37 no.2
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    • pp.227-247
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    • 2013
  • MicroRNAs (miRNAs) are a class of recently discovered non-coding small RNA molecules, on average approximately 21 nucleotides in length, which underlie numerous important biological roles in gene regulation in various organisms. The miRNA database (release 18) has 18,226 miRNAs, which have been deposited from different species. Although miRNAs have been identified and validated in many plant species, no studies have been reported on discovering miRNAs in Panax ginseng Meyer, which is a traditionally known medicinal plant in oriental medicine, also known as Korean ginseng. It has triterpene ginseng saponins called ginsenosides, which are responsible for its various pharmacological activities. Predicting conserved miRNAs by homology-based analysis with available expressed sequence tag (EST) sequences can be powerful, if the species lacks whole genome sequence information. In this study by using the EST based computational approach, 69 conserved miRNAs belonging to 44 miRNA families were identified in Korean ginseng. The digital gene expression patterns of predicted conserved miRNAs were analyzed by deep sequencing using small RNA sequences of flower buds, leaves, and lateral roots. We have found that many of the identified miRNAs showed tissue specific expressions. Using the insilico method, 346 potential targets were identified for the predicted 69 conserved miRNAs by searching the ginseng EST database, and the predicted targets were mainly involved in secondary metabolic processes, responses to biotic and abiotic stress, and transcription regulator activities, as well as a variety of other metabolic processes.