• Title/Summary/Keyword: DNA homology

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Expression of the Galactose Mutarotase Gene from Lactococcus lactis ssp. lactis ATCC7962 in Escherichia coli

  • Lee, Jong-Hoon;Choi, Jae-Yeon;Lee, Jung-Min;Kim, Jeong-Hwan;Chang, Hae-Choon;Chung, Dae-Kyun;Lee, Hyong-Joo
    • Journal of Microbiology and Biotechnology
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    • v.10 no.6
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    • pp.840-843
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    • 2000
  • The structure of gal/lac operon of Lactococcus lactis ssp. lactis ATCC7962 was partially characterized and the gene (galM) encoding galactose mutarotase was cloned together with the order; galA-galM-galK-galT. The galM was found to be 1,027 bp in length and encoded the protein of 37,609 Da calculated molecular mass. The deduced amino acid sequence showed a homology with GalM proteins from several other microorganisms. Thus, the galM gene was expressed in Escherichia coli and the product was identified as a 38 kDa protein which corresponded to the size estimated from DNA sequence. mutarotase activity of the IPTG inducedrecombinant was 2.7 times increased against that of the host strain.

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Cloning and Expression Analysis of Gonadogenesis-associated Gene SPATA4 from Rainbow Trout (Oncorhynchus mykiss)

  • Liu, Bowen;Liu, Shangfeng;He, Shan;Zhao, Ying;Hu, Hongxia;Wang, Zhao
    • BMB Reports
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    • v.38 no.2
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    • pp.206-210
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    • 2005
  • Gonadogenesis is a complicated process which involves multi-gene interactions. A rainbow trout (Oncorhynchus mykiss) gene spermatogenesis associated 4 (SPATA4) was cloned and characterized from adult rainbow trout testis. The cDNA sequence of rainbow trout SPATA4 contains an open reading frame of 1, 081 nucleatides encoding a putative protein of 259 amino acids. The putative protein from rainbow trout shares a 76.8% homology with zebrafish SPATA4. No trans-membrane regions or signal peptide were detected using bioinformatics methods. Subcellular localization analysis revealed that rainbow trout SPATA4 was a nuclear protein with highest possibility (39.1%). Multi-tissue reverse transcriptase PCR (RT-PCR) was performed to examine the distribution of rainbow trout SPATA4 in eleven organs of adult rainbow trout. The result demonstrated that this gene express specifically in testis and slight amount of expression was detected in ovary. Further analysis of SPATA4 characterization and function in rainbow trout may provide insight into the understanding of gonadogenesis process.

Molecular Cloning of the Bombyx Ubiquitin Holmologue Gene That Is Up-regulated Upon Infection

  • Yun, Eun-Young;Goo, Tae-Won;Hwang, Jae-Sam;Kang, Seok-Woo;Park, Soo-Jung;Kwon, O-Yu
    • International Journal of Industrial Entomology and Biomaterials
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    • v.2 no.1
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    • pp.61-64
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    • 2001
  • Ubiquitin can be covalently attached to cellular proteins as a post-translational modification rind is involved in metabolic stresses, such as bent shock and immune response. We have isolated and sequenced a cDNA encoding ubiquitin from the silkworm, Bombyx mori. The insert in the clone is 533 nucleotide long with an open reading frame of 387 nucleotides that encodes a protein of 129 amino acids with a molecular weight of 14.8 kDa. The amino acid sequence shared high homology with the ubiquitins known so far, The result of dot blot hybridization showed that the B. mori ubiquitin gene is up-regulated upon f. rofi infection, suggesting that the B. mori ubiquitin plays an immune-related role.

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Molecular cloning and characterization of peroxiredoxin from Toxoplasma gondii

  • Son, Eui-Sun;Song, Kyoung-Ju;Shin, Jong-Chul;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.39 no.2
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    • pp.133-141
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    • 2001
  • A cDNA of 1.1 kb comprising the gene encoding the peroxiredoxin of Toxo-plasma gondii(TgPrx) has been cloned. The open reading frame of 591 Up was translated into a protein of 196 amino acids with a molecular mass of 25 kDa. Conserved 2 cysteine domains of Phe-Val-Cys-Pro and Glu-Val-Cys-Pro indicated TgPrx belonged to 2-Cys Prx families. TgPrx showed the highest homology with that of Arabidopsis thaliana by 53.9% followed by Entamoeba histolytica with 39.5% by the amino acid sequence alignment. Polyclonal antibody against recombinant TgPrx detected 25 kDa band in T. gondii without binding to host cell proteins TgPrx was located in the cytoplasm of T. gondii extracellularly or intracellularly by immunofluorescence assay. The expression of TgPrx was increased as early as 30 min after the treatment with artemisinin in the intracellular stage, while no changes in those of host Prx I and TgSOD. This result implies that TgPrx may function as an antioxidant protecting the cell from the attack of reactive oxygen intermediates. It is also suggested that TgPrx is a possible target of chemotherapy.

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Painless Jaundice Caused by Clonorchis sinensis Infection: A Case Report

  • Hao, Yuhua;Bao, Wanguo;Jin, Meishan;Li, Yuxiang;Wang, Feng
    • Parasites, Hosts and Diseases
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    • v.54 no.3
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    • pp.323-327
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    • 2016
  • A man with only yellowing of the skin and eye sclera was diagnosed with clonorchiasis, which rarely manifested jaundice as the initial symptom. However, because of a lack of evidence for a diagnostic gold standard, the time until definitive diagnosis was more than a week. The diagnostic process relied on inquiring about the patient's history, including the place of residence, dietary habits, and symptoms, as well as on serological findings, an imaging examination, and pathological findings. MRCP and CT results showed mild dilatation of intrahepatic ducts and increased periductal echogenicity. The eggs were ultimately found in stool by water sedimentation method after the negative report through direct smear. DNA sequencing of PCR production of the eggs demonstrated 98-100% homology with ITS2 of Clonorchis sinensis. After anti-parasite medical treatment, the patient's symptoms were gradually relieved. Throughout the diagnostic procedure, besides routine examinations, the sedimentation method or concentration method could be used as a sensitive way for both light and heavy C. sinensis infection in the definite diagnosis.

Modification of cell wall structural carbohydrate in the hybrid poplar expressing Medicago R2R3-MYB transcription factor MtMYB70

  • Kim, Sun Hee;Choi, Young Im;Jin, Hyunjung;Shin, Soo-Jeong;Park, Jong-Sug;Kwon, Mi
    • Journal of Plant Biotechnology
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    • v.42 no.2
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    • pp.93-103
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    • 2015
  • The isolation, cloning, and characterization of an R2R3-MYB transcription factor gene (MtMYB70) from the model legume Medicago truncatula is reported. MtMYB70 consists of a 768-bp coding sequence corresponding to 255 amino acids. Sequence alignment revealed that MtMYB70 cDNA contains conserved R2R3-type MYB domains with highly divergent C terminal regions. MtMYB70 was found to have relatively low sequence homology with known R2R3-MYB genes. Phylogenetic analysis placed the R2R3-MYB domain of MtMYB70 closest to PtMYB1, a known activator of lignin biosynthesis. Overexpression of MtMYB70 under the control of the 35S promoter in transgenic poplar did not cause a significant difference in total lignin content relative to the control, but glucan content was significantly increased in transgenic poplar. Therefore, MtMYB70 might have regulatory role in the biosynthesis of cell wall structural carbohydrates.

Sll0396 regulates transcription of the phycocyanin genes in Synechocystis sp. PCC 6803

  • Oh, In-Hye;Kim, Ho-San;Chung, Young-Ho;Kim, Young-Hye;Park, Young-Mok
    • Plant Biotechnology Reports
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    • v.4 no.3
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    • pp.193-199
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    • 2010
  • An olive-green mutant was generated in Synechocystis sp. strain PCC 6803 by inactivation of the sll0396 gene. Whole-cell absorption spectra of the mutant revealed the missing of phycocyanin peak. An investigation of the low-temperature fluorescence emission spectra revealed that the $sll0396{\Omega}$ mutant has a reduced amount of phycocyanin. Western blot analysis showed that the mutant contained less phycocyanin ${\beta}$- and ${\alpha}$-subunits and lacked the 30- and 32-kDa linker polypeptides, and northern blot analysis revealed that the transcription of the 1.4-kb cpcBA gene encoding the phycocyanin ${\beta}$- and ${\alpha}$-subunits was lower in the mutant. The Sll0396 protein has a DNA-binding motif and shares homology with known response regulators. Our results indicate that Sll0396 plays a regulatory role in the transcription of the phycocyanin genes during phycobilisome synthesis.

Molecular cloning and expression of glyceraldehyde-3-phosphate dehydrogenase gene under environmental stresses in sweetpotato

  • Kim, Young-Hwa;Song, Young-Sun;Huh, Gyung-Hye
    • Journal of Plant Biotechnology
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    • v.35 no.2
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    • pp.95-100
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    • 2008
  • Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a main enzyme in the glycolytic pathway, is involved in cellular energy production and regarded as a housekeeping gene. Previously, cytosolic GAPDH was selected as the most significantly abundant gene in EST library of sweetpotato suspension cells. In this study, a full-length of cDNA clone (IbGAPDH) encoding GAPDH was isolated from suspension-cultured cells of sweetpotato (Ipomoea babatas), and its expression was investigated with a view to understanding the physiological function of GAPDH in relation to environmental stresses. IbGAPDH encoded a 36.9 kDa polypeptide consisting of 337 amino acids. When the deduced amino acid of IbGAPDH was compared with other higher plants, IbGAPDH showed high homology with cytosolic GAPDH. The mRNA level of IbGAPDH significantly increased under environmental stresses, such as $H_2O_2$, MV and cold treatments. Among them, the transcript level of IbGAPDH gene was the highest under cold stress. Further investigation of the transcription level under $10^{\circ}C$ or $15^{\circ}C$ was performed with different tissues of sweetpotato. The transcription of IbGAPDH was increased by cold stress with tissue-specificity, moreover, showed different patterns according to temperature.

Identification of Culturable Bioaerosols Collected over Dryland in Northwest China: Observation using a Tethered Balloon

  • Chen, Bin;Kobayashi, Fumihisa;Yamada, Maromu;Kim, Yang-Hoon;Iwasaka, Yasunobu;Shi, Guang-Yu
    • Asian Journal of Atmospheric Environment
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    • v.5 no.3
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    • pp.172-180
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    • 2011
  • The transfer of microorganisms is important process for ecosystems. Microorganisms in dryland can transport itself to wetland through atmospheric diffusion, but only few papers reported about the atmospheric bioaerosol present over dryland. We carried out the direct sampling using a tethered balloon over Dunhuang City, China's northwestern dryland. Bioaerosols were collected using a tethered balloon with a bioaerosol collector at 820 m above the ground (1,960 m above the sea level) around noon on August 17, 2007. The bioaerosols were cultured after the collection at Dunhuang Meteorological observatory. Two strains of molds were isolated using the Nutrient agar medium. About 400-bp 18S rRNA partial sequences were amplified by PCR and determined afterwards. The results of a homology search by 18S rRNA sequences of isolates in DNA databases (GenBank, DDBJ, and EMBL) and an observation of the form revealed that two bioaerosols in the convective mixed layer over Dunhuang City were Cladosporium sp. and Aspergillus sp.

Analysis of Expressed Sequence Tags from the Wood-Decaying Fungus Fomitopsis palustris and Identification of Potential Genes Involved in the Decay Process

  • Karim, Nurul;Shibuya, Hajime;Kikuchi, Taisei
    • Journal of Microbiology and Biotechnology
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    • v.21 no.4
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    • pp.347-358
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    • 2011
  • Fomitopsis palustris, a brown-rot basidiomycete, causes the most destructive type of decay in wooden structures. In spite of its great economic importance, very little information is available at the molecular level regarding its complex decay process. To address this, we generated over 3,000 expressed sequence tags (ESTs) from a cDNA library constructed from F. palustris. Clustering of 3,095 high-quality ESTs resulted in a set of 1,403 putative unigenes comprising 485 contigs and 918 singlets. Homology searches based on BlastX analysis revealed that 78% of the F. palustris unigenes had a significant match to proteins deposited in the nonredundant databases. A subset of F. palustris unigenes showed similarity to the carbohydrateactive enzymes (CAZymes), including a range of glycosyl hydrolase (GH) family proteins. Some of these CAZyme-encoded genes were previously undescribed for F. palustris but predicted to have potential roles in biodegradation of wood. Among them, we identified and characterized a gene (FpCel45A) encoding the GH family 45 endoglucanase. Moreover, we also provided functional classification of 473 (34%) of F. palustris unigenes using the Gene Ontology hierarchy. The annotated EST data sets and related analysis may be useful in providing an initial insight into the genetic background of F. palustris.