• Title/Summary/Keyword: C-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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Comparison Between Phylogenetic Relationships Based on 18S rDNA Sequences and Growth by Salinity of Chlorella-like Species (Chlorophyta)

  • Lee, Hye-Jung;Hur, Sung-Bum
    • Fisheries and Aquatic Sciences
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    • v.15 no.2
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    • pp.125-135
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    • 2012
  • This study was carried out to understand the correlation between phylogenetic relationships based on 18S rDNA sequences and growth by salinity of Chlorella-like species. The 18S rDNA sequences of 71 Chlorella-like species which were mainly collected from Korean waters were analyzed. The 18S rDNA sequences of Chlorella-like species were divided into three groups (group A, B and C) and group B was further divided into three subgroups (subgroup B-1, B-2 and B-3). Thirty-seven Chlorella-like species in group A grew well at high salinity (32 psu) but the other groups grew well in freshwater. The sequence identities of the species in group A and B were 97.2-99.5%, but those of 6 species in group C ("Chlorella" saccharophila), which contained group I intron sequences region were 75.0-75.4%. Two representative species of each group were cultured at different salinities (0, 16 and 32 psu) to examine the correlation between the molecular phylogenetic groups and the phenotypic characteristics on cell growth and size by different salinities. The size of cell cultured at different salinities varied according to the species of each molecular phylogenetic group. The size of "Chlorella" saccharophila in group C was bigger and more obviously elliptical rather than that of the other Chlorella-like species. Considering the results on molecular and phenotypic characteristics, the group A and B belonged to Chlorellaceae, but group C was distinctly different from them.

Genogroup position of aquabirnavirus GC-1 isolated from rockfish Sebastes schiegeli in Korea

  • Joh, Seong-Joon;Lee, Youn-Jeong;Song, Chang-Sun;Kang, Shien-Young;Mo, In-Pil;Heo, Gang-Jun
    • Korean Journal of Veterinary Research
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    • v.48 no.3
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    • pp.287-293
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    • 2008
  • The cDNA of the aquabirnavirus, GC-1 isolated from rockfish Sebastes schlegeli in Korea, was synthesized using the reverse transcriptase-polymerase chain reaction. The nucleotide and deduced amino acid sequences were determined from cDNA of the VP2-NS-VP3 coding region of genome segment A. The nucleotide sequences of the segment A were 3,086 base pairs (bp) in length and contained large open reading frame (ORF) and terminal sequences. The large ORF was comprised of 2,916 bp nucleotides and composed of 972 deduced amino acid sequences. Pairwise comparisons were made with other aquabirnavirus sequences published previously. The study of genetic relationships between GC-1 and aquabirnaviruses in the large ORF and VP2 coding regions demonstrated that the GC-1 has the nearest genetic relationship with the marine birnaviruses (MABV strains), and the GC-1 and MABV strains can be clustered as the same genogroup. GC-1 can be included in MABV, which is the 7th genogroup of family Aquabirnaviridae.

Molecular Cloning of Chicken Major Histocompatibility Complex Class II Molecules

  • Sung, Aree-Moon
    • Toxicological Research
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    • v.8 no.2
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    • pp.331-342
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    • 1992
  • The chicken major histocompatibility complex (MHC), the B complex, is beginning to be analyzed at the DNA level. Inbred lines of chickens have been reported to possess 3~5 MHC class II genes. To further analyzed the molecular structure of the chicken MHC class II genes, cDNA clones coding for chicken MHC class II (B-L) ${\beta}$ chain molecules were isolated from chicken spleen and liver. Tissue-specific transcription of B-L ${\beta}$genes was studied by Northern blot analysis. A high level of expression was detected for spleen poly(A)$^+$ RNA whereas a faint signal was detected for liver poly(A)$^+$ RNA. Twenty-nine cDNA clones were isolated from the spleen and eight cDNA clones were isolated from the liver. Based on restriction maps, most clones could be clustered into one family of genes. Four cDNA clones were sequenced (S7, S10 and S19 from the spleen and L1, which was identical to S19, from the liver). Complete amino acid sequences of B-L ${\beta}$ chain molecules were predicated from the nucleotide sequences of the cDNA clones. Although both the nature and the location of the conserved residues were similar in chicken and mammalian sequences, some species-specific differences were found, suggesting that the structures of the B-L molecules are similar, but not identical to their mammalian counterparts.

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Phylogenetic Relationship Among Four Species of Korean Oysters Based on Mitochondrial 16S rDNA and COI Gene (미토콘드리아 16S rDNA와 COI유전자에 근거한 한국산 굴류 4종의 유연관계)

  • 이상엽;박두원;안혜숙;김상해
    • Animal Systematics, Evolution and Diversity
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    • v.16 no.2
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    • pp.203-211
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    • 2000
  • Partial mitochondrial 16S rDNA and COI gene were amplified using PCR and sequenced for four species of oysters in Korea. Phylogenetic relationships among them were inferred from their aligned sequences by neighbor-joining method. The sequence comparison data of two mitochondrial genes showed that the genetic distinction between two oyster genera (Crassostreo and Ostrea) was obvious. Phylogenetic analysis based on the nucleotide sequences and A+T percentage of two genes indicates that C. gigas and C. nippona strongly formed a sister group and then C. ariakensis was clustered with the clade although that based on amino acid sequences of COI gene by neighbor-joining method represented different phylogenetic tree.

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Sequencing of cDNA Clones Expressed in Adipose Tissues of Korean Cattle

  • Bong, J.J.;Tong, K.;Cho, K.K.;Baik, M.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.4
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    • pp.483-489
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    • 2005
  • To understand the molecular mechanisms that regulate intramuscular fat deposition and its release, cDNA clones expressed in adipose tissues of Korean cattle were identified by differential screening from adipose tissue cDNA library. By partial nucleotide sequencing of 486 clones and a search for sequence similarity in NCBI nucleotide databases, 245 clones revealed unique clones. By a functional grouping of the clones, 14% of the clones were categorized to metabolism and enzyme-related group (stearoyl CoA desaturase, lactate dehydrogenase, fatty acid synthase, ATP citrate lyase, lipoprotein lipase, acetyl CoA synthetase, etc), and 6% to signal transduction/cell cycle-related group (C/EBP, cAMP-regulated phosphoprotein, calmodulin, cyclin G1, cyclin H, etc), and 4% to cytoskeleton and extracellular matrix components (vimentin, ankyrin 2, gelosin, syntenin, talin, prefoldin 5). The obtained 245 clones will be useful to study lipid metabolism and signal transduction pathway in adipose tissues and to study obesity in human. Some clones were subjected to full-sequencing containing open reading frame. The cDNA clone of bovine homolog of human prefoldin 5 gene had a total length of 959 nucleotides coding for 139 amino acids. Comparison of the deduced amino acid sequences of bovine prefoldin 5 with those of human and mouse showed over 95% identity. The cDNA clone of bovine homolog of human ubiquitin-like/S30 ribosomal fusion protein gene had a total length of 484 nucleotides coding for 133 amino acids. Comparison of the deduced amino acid sequences of bovine ubiquitin-like/S30 ribosomal fusion protein gene with those of human, rat and mouse showed over 97% identity. The cDNA clone of bovine homolog of human proteolipid protein 2 mRNA had a total length of 928 nucleotides coding for 152 amino acids. Comparison of the deduced amino acid sequences of bovine proteolipid protein 2 with those of human and mouse showed 87.5% similarity. The cDNA clone of bovine homolog of rat thymosin beta 4 had a total length of 602 nucleotides coding for 44 amino acids. Comparison of the deduced amino acid sequences of bovine thymosin beta 4 gene with those of human, mouse and rat showed 93.1% similarity. The cDNA clone of bovine homolog of human myotrophin mRNA had a total length of 790 nucleotides coding for 118 amino acids. Comparison of the deduced amino acid sequences of bovine myotrophin gene with those of human, mouse and rat showed 83.9% similarity. The functional role of these clones in adipose tissues needs to be established.

Polyphase Signature Sequences for M-ary Phase Signaling (M진 위상 신호 방식에 효과적인 다상 서명 수열)

  • Park, So-Ryoung;Song, Iick-Ho
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.32 no.11C
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    • pp.1059-1065
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    • 2007
  • In this paper, we propose a class of polyphase signature sequences, whose general odd correlation properties are useful for M-ary phase signaling systems. The maximum magnitude of the general odd correlation functions of the proposed sequences are investigated and compared with those of FZC (Frank-Zadoff-Chu) sequences and those of EOE (equivalent odd and even correlation) sequences. The performance of the asynchronous M-ary phase signaling systems using the proposed sequences is simulated and compared with that using FZC sequences in direct sequence code division multiple access (DS/CDMA) systems. The performance of the system using the proposed sequence is shown to be better than that using FZC sequences when the multipath fading is in existence.

Co-expression of IRES-mediated hG-CSF cDNA and hGH Gene under the Control of Goat beta-Casein Promoter

  • Oh, Keon-Bong;Lee, Chul-Sang
    • Development and Reproduction
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    • v.14 no.1
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    • pp.13-19
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    • 2010
  • We developed a novel dicistronic system for the expression of target cDNA sequences in the milk of transgenic animals using goat beta-casein/hGH fusion construct, pGbc5.5hGH (Lee, 2006) and internal ribosome entry site (IRES) sequences of encephalomyocarditis virus (EMCV). Granulocyte colony-stimulating factor (hG-CSF) cDNA was linked to 3' untranslated region of hGH gene in the pGbc5.5hGH via EMCV IRES sequences. Transgenic mice were generated by microinjection and transgene expression was examined in the milk and mammary gland of transgenic mice at 10 days of lactation. Northern blot analysis showed that hGH gene and hG-CSF cDNA were transcribed as a single dicistronic mRNA. The hG-CSF and hGH proteins were independently translated from the dicistronic mRNA and secreted into the milk of transgenic mice. The highest concentration of hG-CSF and hGH in the milk of transgenic mice were $237{\mu}g/m{\ell}$ and $8,990{\mu}g/m{\ell}$, respectively. In contrast, another hG-CSF expression cassette, in which hG-CSF genomic sequences were inserted into a commercial milk-specific expression vector (pBC1), generated a lower level ($91{\mu}g/m{\ell}$) of hG-CSF expression in the milk of transgenic mice. These results demonstrated that the novel pGbc5.5hGH-based dicistronic construct could be useful for an efficient cDNA expression in the milk of transgenic animals.

Analyses of Expressed Sequence Tags from Chironomus riparius Using Pyrosequencing : Molecular Ecotoxicology Perspective

  • Nair, Prakash M. Gopalakrishnan;Park, Sun-Young;Choi, Jin-Hee
    • Environmental Analysis Health and Toxicology
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    • v.26
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    • pp.10.1-10.7
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    • 2011
  • Objects: Chironomus riparius, a non-biting midge (Chironomidae, Diptera), is extensively used as a model organism in aquatic ecotoxicological studies, and considering the potential of C. riparius larvae as a bio-monitoring species, little is known about its genome sequences. This study reports the results of an Expressed Sequence Tags (ESTs) sequencing project conducted on C. riparius larvae using 454 pyrosequencing. Method: To gain a better understanding of C. riparius transcriptome, we generated ESTs database of C.ripairus using pyrosequencing method. Results: Sequencing runs, using normalized cDNA collections from fourth instar larvae, yielded 20,020 expressed sequence tags, which were assembled into 8,565 contigs and 11,455 singletons. Sequence analysis was performed by BlastX search against the National Center for Biotechnology Information (NCBI) nucleotide (nr) and uniprot protein database. Based on the gene ontology classifications, 24% (E-value${\leq}1^{-5}$) of the sequences had known gene functions, 24% had unknown functions and 52% of sequences did not match any known sequences in the existing database. Sequence comparison revealed 81% of the genes have homologous genes among other insects belonging to the order Diptera providing tools for comparative genome analyses. Targeted searches using these annotations identified genes associated with essential metabolic pathways, signaling pathways, detoxification of toxic metabolites and stress response genes of ecotoxicological interest. Conclusions: The results obtained from this study would eventually make ecotoxicogenomics possible in a truly environmentally relevant species, such as, C. riparius.

Gene Expression Profiling of Eukaryotic Microalga, Haematococcus pluvialis

  • EOM HYUNSUK;PARK SEUNGHYE;LEE CHOUL-GYUN;JIN EONSEON
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.1060-1066
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    • 2005
  • Under environmental stress, such as strong irradiance or nitrogen deficiency, unicellular green algae of the genus Haematococcus accumulate secondary carotenoids, i.e. astaxanthin, in the cytosol. The induction and regulation of astaxanthin biosynthesis in microalgae has recently received considerable attention owing to the increasing use of secondary carotenoids as a source of pigmentation for fish aquacultures, and as a potential drug in cancer prevention as a free-radical quencher. Accordingly, this study generated expressed sequence tags (ESTs) from a library constructed from astaxanthin-induced Haematococcus pluvialis. Partial sequences were obtained from the 5' ends of 1,858 individual cDNAs, and then grouped into 1,025 non-overlapping sequences, among which 708 sequences were singletons, while the remainder fell into 317 clusters. Approximately $63\%$ of the EST sequences showed similarity to previously described sequences in public databases. H. pluvialis was found to consist of a relatively high percentage of genes involved in genetic information processing ($15\%$) and metabolism ($11\%$), whereas a relatively low percentage of sequences was involved in the signal transduction ($3\%$), structure ($2\%$), and environmental information process ($3\%$). In addition, a relatively large fraction of H. pluvialis sequences was classified as genes involved in photosynthesis ($9\%$) and cellular process ($9\%$). Based on this EST analysis, the full-length cDNA sequence for superoxide dismutase (SOD) of H. pluvialis was cloned, and the expression of this gene was investigated. The abundance of SOD changed substantially in response to different culture conditions, indicating the possible regulation of this gene in H. pluvialis.