• Title/Summary/Keyword: conserved sequences

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MOLECULAR CLONING OF CHICKEN INTERFERON-GAMMA (닭 인터페론 유전자의 클로닝에 관한 연구)

  • ;Hyun Lillehoj
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 1999.11a
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    • pp.34-50
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    • 1999
  • A cDNA encoding chicken interferon-gamma (chIFN-${\gamma}$) was amplified from P34, a CD4$^{+}$ T-cell hybridoma by reverse transcription-polymerase chain reaction (RT-PCR) and cloned into pUC18. THe sequences of cloned PCR products were determined to confirm the correct cloning. Using this cDNA as probe, chicken genomic library from White Leghorn spleen was screened. Phage clones harboring chicken interferon-gamma (chIFN-${\gamma}$) were isolated and their genomic structure elucidated. The chIFN-${\gamma}$ contains 4 exons and 3 introns spanning over 14 kb, and follows the GT/AG rule for correct splicing at the exon/intron boundaries. The four exons encode 41, 26, 57 and 40 amino acids, respectively, suggesting that the overall structure of IFN-${\gamma}$ is evolutionairly conserved in mammalian and avian species. The 5’-untranslated region and signal sequences are located in exon 1. Several AT-rich sequences located in the fourth exon may indicate a role in mRNA turnover. The 5’-flanking region contains sequences homologous to the potential binding sites for the mammalian transcription factors, activator protein-1(AP-1) activator protein-2(AP-2) cAMP-response element binding protein(CREB), activating transcription factor(ATF), GATA-binding fator(GATA), upstream stimulating factor(USF), This suggests that the mechanisms underlying transcriptional regulation of chicken and mammalian IFN-${\gamma}$ genes may be similar.r.

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First Report on the Witches' Broom in Annual Statice (Limonium sinuatum) in Korea

  • Chung, Bong-Nam;Huh, Kun-Yang;Jeong, Myeong-Il
    • The Plant Pathology Journal
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    • v.21 no.4
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    • pp.383-386
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    • 2005
  • In 2003 typical phytoplasma symptoms of witches' broom and flower malformation were observed on statice (Limonium sinuatum) plants grown at commercial greenhouses in Busan, South Korea. The DNA extracted from the infected leaves was amplified using universal primer pair of Pl/P6 derived from conserved 16S rRNA gene of Mollicutes giving the expected Polymerase chain reaction (PCR) product of 1.5 kb. In the nested PCR assays, the expected DNA fragment of 1.1 kb was amplified with the specific primer pair 16Fl/Rl that was designed on the basis of aster yellows (AY) phytoplasma 16S rDNA sequences. The 1.1 kb PCR products were cloned and nucleotide sequences were determined. The sequences were identical to that of Onion yellows OY phytoplasma (GenBank accession no. D12569) isolated from Onion in Japan. Electron microscopy of thin sections of leaf veins showed phytoplasma bodies in the phloem. Statice witches' broom symptom occurred on statice in commercial greenhouses in Korea was confirmed as infection of AY phytoplasma by transmission electron microscopy observation, and by determination of 16S rRNA gene sequences of phytoplasma.

Sturcture of the Rice Glutelin Multigene Family and Its Expression (쌀 Glutelin 유전자군의 구조 및 발현조절)

  • 황영수
    • Proceedings of the Botanical Society of Korea Conference
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    • 1987.07a
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    • pp.261-282
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    • 1987
  • Plants store a significant amount of their nitrogen, sulfur and carbon reserves as storage proteins in seed tissues. The major proteins present in rice seeds are the glutelins. Glutelins are initially synthesized at 4-6 days postanthesis and deposited into protein bodies via Golgi apparatus. Based on nucleic acid sequences and Southern blot analysis, the three isolated glutelin genomic clones were representative members of three gene subfamilies each containing 5 to 8 copies. A comparison of DNA sequences displayed by relevant regions of these genomic clones showed that two subfamilies, represented by clones, Gt1 and Gt2, were closely, related and probably evolved by more recent gene duplication events. The 5' flanking and coding sequences of Gt1 and Gt2 displayed at least 87% homolgy. In contrast, Gt3 showed little or no homolgy in the 5' flanking sequences upstream of the putative CAAT boxes and exhibited significant divergence in all other portions of the gene. Conserved sequences in the 5' flanking regions of these genes were identified and discussed in light of their potential regulatory role. The derived primary sequences of all three glutelin genomic clones showed significant homology to the legume 11S storage proteins indicating a common gene origin. A comparison of the derived glutelin primary sequences showed that mutations were clustered in three peptide regions. One peptide region corresponded to the highly rautable hypervariable region of legume peptide region of legume 11S storage proteins, a potential target area for protein modification. Expression studies indicated that glutelin mRNA transcripts are differentially accumulated during endosperm development. Promoterss of Gt2 and Gt3 were functional as they direct transient expression of chloramphenicol acetyltransferase in cultured plant cell.

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Site-directed Mutagenesis of Five Conserved Residues of Subunit I of the Cytochrome cbb3 Oxidase in Rhodobacter capsulatus

  • Ozturk, Mehmet;Gurel, Ekrem;Watmough, Nicholas J.;Mandaci, Sevnur
    • BMB Reports
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    • v.40 no.5
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    • pp.697-707
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    • 2007
  • Cytochrome $cbb_3$ oxidase is a member of the heme-copper oxidase superfamily that catalyses the reduction of molecular oxygen to the water and conserves the liberated energy in the form of a proton gradient. Comparison of the amino acid sequences of subunit I from different classes of heme-copper oxidases showed that transmembrane helix VIII and the loop between transmembrane helices IX and X contain five highly conserved polar residues; Ser333, Ser340, Thr350, Asn390 and Thr394. To determine the relationship between these conserved amino acids and the activity and assembly of the $cbb_3$ oxidase in Rhodobacter capsulatus, each of these five conserved amino acids was substituted for alanine by site-directed mutagenesis. The effects of these mutations on catalytic activity were determined using a NADI plate assay and by measurements of the rate of oxygen consumption. The consequence of these mutations for the structural integrity of the $cbb_3$ oxidase was determined by SDS-PAGE analysis of chromatophore membranes followed by TMBZ staining. The results indicate that the Asn390Ala mutation led to a complete loss of enzyme activity and that the Ser333Ala mutation decreased the activity significantly. The remaining mutants cause a partial loss of catalytic activity. All of the mutant enzymes, except Asn390Ala, were apparently correctly assembled and stable in the membrane of the R. capsulatus.

Cloning, Sequencing and Expression Analysis of Porcine Uroplakin II Gene

  • Gwon Deuk-Nam;Kim Jin-Hoe
    • Proceedings of the KSAR Conference
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    • 2002.06a
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    • pp.90-90
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    • 2002
  • In this study, we report the cloning of the porcine UPII genomic DNA, which contains a putative full-length open reading frame encoding the UPII protein. A comparison of the porcine UPII gene coding sequence with the previously published mouse UPII sequence demonstrates that only the exon sequences are partially conserved. Northern and immunohistochemical analyses show that the porcine UPII gene is expressed only in the urothelium and that the protein specifically localizes to urothelial superficial cells. (omitted)

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Isolation and Characterization of Parvalbumin Beta Gene from Channel Catfish (Ictalurus punctatus)

  • Kim, Soon-Hag
    • Journal of Aquaculture
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    • v.16 no.2
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    • pp.124-127
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    • 2003
  • Our previous studies of both microarray analysis in channel catfish muscle gene expression of 2 different ages and channel catfish muscle expressed sequence tag profiles demonstrated parvalbumin beta is one of the highly expressed muscle transcriptome. We have cloned and sequenced complementary DNA encoding the channel catfish parvalbumin which encode 109 amino acids. The deduced amino acid sequences of the catfish parvalbumin are highly conserved with those cloned from other teleosts. The availability of the catfish parvalbumin provides the opportunity of studying fish epitopes.

SECONDARY STRUCTURE OF THE PANHANDLE RNA OF INFLUENZA VIRUS A STUDIED BY NMR SPECTROSCOPY

  • Cheong, Hae-Kap;Park, Byong-Seok;Chaejoon Cheong
    • Proceedings of the Korean Biophysical Society Conference
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    • 1996.07a
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    • pp.31-31
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    • 1996
  • The double-stranded panhandle structure of the influenza virus RNA is important for the replication, transcription and packaging into the virion of the vRNA. The solution structure of a 34-nucleotide-long RNA which contains the conserved panhandle sequences has been investigated by one- and two-dimensional NMR spectroscopies. (omitted)

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In Silico Metagenomes Mining to Discover Novel Esterases with Industrial Application by Sequential Search Strategies

  • Barriuso, Jorge;Jesus Martinez, Maria
    • Journal of Microbiology and Biotechnology
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    • v.25 no.5
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    • pp.732-737
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    • 2015
  • We present here an in silico search of fungal sterol-esterase/lipase and bacterial depolymerase sequences from environmental metagenomes. Both enzyme types contain the α/β-hydrolase protein fold. Analysis of DNA conserved motifs, protein homology search, phylogenetic analysis, and protein 3D modeling have been used, and the efficiency of these screening strategies is discussed. The presence of bacterial genes in the metagenomes was higher than those from fungi, and the sequencing depth of the metagenomes seemed to be crucial to allow finding enough diversity of enzyme sequences. As a result, a novel putative PHA-depolymerase is described.

Plant RNA Virus-Host Interaction: Potato virus X as a model system

  • Kim, Kook-Hyung
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.14-14
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    • 2003
  • Potato virus X (PVX), the type member of Potexvirus genus, is a flexuous rod-shaped virus containing a single-stranded (+) RNA. Infection by PVX produces genomic plus- and minus-strand RNAs and two major subgenomic RNAs (sgRNAs). To understand the mechanism for PVX replication, we are studying the cis- and/or trans-acting elements required for RNA replication. Previous studies have shown that the conserved sequences located upstream of two major sgRNAs, as well as elements in the 5' non-translated region (NTR) affect accumulation of genomic and sg RNAs. Complementarity between sequences at the 5' NTR and those located upstream of two major sgRNAs and the binding of host protein(s) to the 5' NTR have shown to be important for PVX RNA replication. The 5 NTR of PVX contains single-stranded AC-rich sequence and stem-loop structure. The potential role(s) of these cis-elements on virus replication, assembly, and their interaction with viral and host protein(s) during virus infection will be discussed based on the data obtained by in vitro binding, in vitro assembly, gel shift mobility assay, host gene expression profiling using various mutants at these regions.

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A Probabilistic Approach to the Assessment of Phylogenetic Conservation in Mammalian Hox Gene Clusters

  • Stojanovic, Nikola;Dewar, Ken
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2005.09a
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    • pp.118-123
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    • 2005
  • With the increasing availability of mammalian genome sequences it became possible to use large scale phylogenetic analysis in order to locate potentially functional regions. In this paper we describe a new probabilistic method for the characterization of phylogenetic conservation in mammalian DNA sequences. We have used this method for the analysis of Hox gene clusters, based on the alignment of 6 species, and we constructed a map of for indicating short and long conserved fragments and their positions with respect to the known locations of Hox genes and other elements, sometimes showing surprising layouts.

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