• Title/Summary/Keyword: DNA Coding

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Cell Cycle Regulated Expression of Subcloned Chicken H3 Histone Genes and Their 5' Flanking Sequences

  • Son, Seung-Yeol;Tae, Gun-Sik
    • Journal of Microbiology and Biotechnology
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    • v.4 no.4
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    • pp.274-277
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    • 1994
  • We subcloned two chicken H3 histone genes and transfected them into Rat 3 cell line. One contains 300 bp 5' to its cap site and the other contains 130 bp 5' to its cap site when cloned into plasm ids. Both of them showed 5' phase specific expression of their mRNA about 8 fold higher (during 5' phase) than during Gl phase. This means that only 130 bp 5' to its cap site was enough to confer cell cycle regulated expression of the latter gene. The DNA sequences of their 5' flanking region did not reveal any particular homologies or subtype-specific sequences. The DNA sequence data also showed that even though the protein coding regions of the histone genes have been conserved exceptionally well throughout evolution, their 5' untranslated regions have not been conserved as well.

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Complete Genome Sequence of Levilactobacillus brevis KL251 Isolate from Kimchi

  • Kiyeop Kim;Da Jeong Shin;Junghee Lee;Sejong Oh
    • Journal of Dairy Science and Biotechnology
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    • v.42 no.1
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    • pp.18-22
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    • 2024
  • In this study, we performed whole-genome sequencing of Levilactobacillus brevis KL251 (KL251) isolated from kimchi. The KL251 genome, characterized by a circular chromosome spanning 2,345,062 base pairs with a GC content of 45.78%, was analyzed. KL251 contains 2,275 coding DNA sequences (CDSs), 56 transfer RNAs (tRNAs), and 4 ribosomal RNAs (rRNAs). Genes associated with gamma-aminobutyric acid (GABA) production and CRISPR-Cas systems were identified and could potentially be used for GABA synthesis and defense against foreign DNA. Additionally, the presence of functional genes involved in isoprenoid biosynthesis, glutathione generation, and redox sensing showed that cellular metabolism and stress responses were important characteristics of this genome. These genomic findings suggest that the KL251 strain could potentially have several applications, including food fermentation, probiotics, dairy product starters, and the development of health-enhancing products.

Sequence diversity of Mitochondrial DNA HV1 in Korean population (한국인 집단의 미토콘드리아 DNA HV1 부위에서의 염기서열 다양성)

  • Lim, Si-Keun;Kim, Eung-Su;Kim, Soon-Hee;Park, Ki-Won;Han, Myun-Soo
    • Analytical Science and Technology
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    • v.18 no.4
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    • pp.362-367
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    • 2005
  • The human mitochondrial genome (mtDNA) has been an important tool in the field of forensic investigations. Within the entire mtDNA molecule, the non-coding control region which is approximately 1,100 bp including hypervariable region I and II (HV1 and HV2) is widely studied because it is highly polymorphic and useful for human identification purposes. In this study, 360 unrelated Koreans were analyzed in HV1. The number of polymorphic sites and genetic lineage were 124 and 210, respectively. The most prevalent substitution was C-T and 75.8% of DNA showed C-T substitution at 16223. There were 20 kinds of polymorphism between 16180 and 16193 including insertion and deletion. The most frequent haplotype was [16223T, 16362C] representing 5%. Approximately 25.9% of DNA showed the same haplotype in at least two samples. The gene diversity was calculated to 0.996 and the probability of two unrelated perosons having the same haplotype was determined to 0.7%.

Complete DNA Sequence and Analysis of a Cryptic Plasmid Isolated from Lactobacillus bifermentans in Kimchi

  • Jeon, Deok-Young;Lee, Sun-Ho
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.1018-1020
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    • 2003
  • The complete 1,486 nucleotide sequence of a cryptic plasmid separated from Lactobacillus bifermentans strain A02 isolated from Kimchi has been determined. The plasmid, designated as pA021, encodes a 33,488 Da putative Rep protein. Based on the sequence similarity, the protein shows homology with coding protein of pRS1, a previously reported plasmid of Oenococcus oeni and the replication initiation protein (Rep) of the Staphylococcal pT181 plasmid family.

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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Complete Mitochondrial Genome of Echinostoma hortense (Digenea: Echinostomatidae)

  • Liu, Ze-Xuan;Zhang, Yan;Liu, Yu-Ting;Chang, Qiao-Cheng;Su, Xin;Fu, Xue;Yue, Dong-Mei;Gao, Yuan;Wang, Chun-Ren
    • Parasites, Hosts and Diseases
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    • v.54 no.2
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    • pp.173-179
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    • 2016
  • Echinostoma hortense (Digenea: Echinostomatidae) is one of the intestinal flukes with medical importance in humans. However, the mitochondrial (mt) genome of this fluke has not been known yet. The present study has determined the complete mt genome sequences of E. hortense and assessed the phylogenetic relationships with other digenean species for which the complete mt genome sequences are available in GenBank using concatenated amino acid sequences inferred from 12 protein-coding genes. The mt genome of E. hortense contained 12 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes, and 1 non-coding region. The length of the mt genome of E. hortense was 14,994 bp, which was somewhat smaller than those of other trematode species. Phylogenetic analyses based on concatenated nucleotide sequence datasets for all 12 protein-coding genes using maximum parsimony (MP) method showed that E. hortense and Hypoderaeum conoideum gathered together, and they were closer to each other than to Fasciolidae and other echinostomatid trematodes. The availability of the complete mt genome sequences of E. hortense provides important genetic markers for diagnostics, population genetics, and evolutionary studies of digeneans.

Transcriptional Activator Elements for Curtovirus C1 Expression Reside in the 3' Coding Region of ORF C1

  • Hur, Jingyung;Buckley, Kenneth J.;Lee, Sukchan;Davis, Keith R.
    • Molecules and Cells
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    • v.23 no.1
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    • pp.80-87
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    • 2007
  • Beet curly top virus (BCTV) and Beet severe curly top virus (BSCTV), members of curtoviruses, encode seven open reading frames (ORFs) within a ~3 kb genome. One of these viral ORFs, C1, is known to play an important role in the early stage of viral infection in plants during initiation of viral DNA replication. We used promoter:: reporter (${\beta}$-glucuronidase) gene fusions in transgenic Arabidopsis to identify the putative promoter region of BCTV ORF C1. Unlike other geminiviruses, the intergenic region of BCTV was not sufficient to promote C1 expression in transgenic plants. When sequences extending into the coding region of C1 were tested, strong expression of the reporter protein was observed in vascular tissues of transgenic plants. This expression was not dependent on the presence of the intergenic regions or proximal 5' portions of the C1 coding region. Transgenic plants expressing a reporter gene under control of the putative complete C1 promoter were inoculated with virus to determine if any viral transcript affected C1 expression. Virus inoculated plants did not show any altered pattern or change in of reporter gene expression level. These results suggest that (1) important transcriptional activator elements for C1 expression reside in the 3' portion of C1 coding area itself, (2) C1 protein does not auto-regulate its own expression and (3) C1 expression of two curtoviruses is controlled differently compared to other geminiviruses.

Transcription and Export of RNase MRP RNA in Xenopus Iaevis Oocyetes

  • Jeong, Seon-Ju
    • Animal cells and systems
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    • v.1 no.2
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    • pp.363-370
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    • 1997
  • RNase MRP is a ribonucleoprotein complex with a site-specific endonuclease activity. Its original substrate for cleavage is the small mitochondrial RNA near the mitochondrial DNA replication origin, thus it was proposed to generate the primer for mtDNA replication. Recently, it has been shown to have another substrate in the nucleus, such as pre-S.8S ribosomal RNA in nucleolus. The gene for the RNA component of RNase MRP (MRP RNA) was found to be encoded by the nucleus genome, suggesting an interesting intracellular trafficking of MRP RNA to both mitochondria and nucleolus after transcription in nucleus. In this study, genomic DNA encoding MRP RNA was microinjected into the nucleus of Xenopus oocytes, to analyze promoter regions involved in the transcription. It showed that the proximal sequence element and TATA box are important for basal level transcription; octamer motif and Sp1 binding sites are for elevated level transcription. Most of Xenopus MRP RNA was exported out to the cytoplasm following transcription in the nucleus. Utilizing various hybrid constructs, export of MRP RNA was found to be regulated by the promoter and the 5' half of the coding region of the gene. Interestingly, the transcription in nucleus seems to be coupled to the export of MRP RNA to cytoplasm. Intracellular transport of injected MRP RNA can be easily visualized by whole-mount in situ hybridization following microinjection; it also shows possible intra-nuclear sites for transcription and export of MRP RNA.

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Cloning and Sequence Analysis of the Cellobiohydrolase I Genes from Some Basidiomycetes

  • Chukeatirote, Ekachai;Maharachchikumbura, Sajeewa S.N.;Wongkham, Shannaphimon;Sysouphanthong, Phongeun;Phookamsak, Rungtiwa;Hyde, Kevin D.
    • Mycobiology
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    • v.40 no.2
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    • pp.107-110
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    • 2012
  • Genes encoding the cellobiohydrolase enzyme (CBHI), designated as cbhI, were isolated from the basidiomycetes Auricularia fuscosuccinea, Pleurotus giganteus, P. eryngii, P. ostreatus, and P. sajor-caju. Initially, the fungal genomic DNA was extracted using a modified cetyltrimethyl ammonium bromide (CTAB) protocol and used as a DNA template. The cbhI genes were then amplified and cloned using the pGEM-T Easy Vector Systems. The sizes of these PCR amplicons were between 700~800 bp. The DNA sequences obtained were similar showing high identity to the cbhI gene family. These cbhI genes were partial consisting of three coding regions and two introns. The deduced amino acid sequences exhibited significant similarity to those of fungal CBHI enzymes belonging to glycosyl hydrolase family 7.

In Vitro Expression of the Recombinant hFSH Gene using Retrovirus Vector System (In Vitro에서 Retrovirus Vector System을 이용한 재조합 hFSH 유전자의 발현)

  • Min, Gyeong-Heon;Kwon, Mo-Sun;Kim, Teoan;Koo, Bon-Chul
    • Reproductive and Developmental Biology
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    • v.35 no.1
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    • pp.115-121
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    • 2011
  • hFSH is a glycoprotein secreted from anterior pituitary and consists of ${\alpha}$ and ${\beta}$ subunits. Because of its major biological functions including sperm formation in the male and for follicular growth, FSH is used to cure woman's sterility. In this study we tried to produce recombinant hFSH in vitro using a retrovirus expression vector. Two major components of the vector we constructed are: ( i ) a DNA fragment containing ${\alpha}$ and ${\beta}$ genes fused by a DNA sequence coding carboxyl terminal peptide (CTP) of human chorionic gonadotropin, (ii) a DNA fragment corresponding woodchuck hepatitis virus posttranscriptional regulatory element (WPRE). Evaluation of expression profile of the recombinant FSH using reverse transcription PCR and enzyme-linked immunosorbent assay (ELISA). Among three cell lines tested, HeLa cells were the best for hFSH expression (5,395 mIU/ml), then followed by chicken embryonic fibroblast (CEF) cells and Chinese hamster ovary (CHO) cells in the order of hFSH production. In addition to the amount, the FSH produced from HeLa cells was highest in terms of biological activity which was determined by measuring cAMP.