• Journal of Microbiology and Biotechnology
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• v.8 no.3
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• pp.214-221
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• 1998

A Brevibacterium ammoniagenes gene coding for glucose/mannose-specific enzyme II ($EII^{Glc}$) of the phosphoenolpyruvate-dependent phosphotransferase system (PTS) was cloned by complementing an Escherichia coli mutation affecting a ptsG gene, and the complete DNA nucleotide sequence was determined. The cloned gene was identified to be a ptsG, which enables the E. coli transportment to use glucose more efficiently than mannose as the sole carbon source in an M9 minimal medium. The ptsG gene of B. ammoniagenes consists of an open reading frame of 1,983 nucleotides putatively encoding a polypeptide of 661 amino acid residues and a TAA stop codon. The deduced amino acid sequence of the B. ammoniagenes $EII^{Glc}$ shows, at $46\%$, the highest degree of sequence similarity with the Corynebacterium glutamicum EII specific for both glucose and mannose. In addition, the $EII^{Glc}$ shares approximately $30\%$ sequence similarities with sucrose-specific and ${\beta}$-glucoside-specific EIIs of the several bacteria belonging to the glucose-PTS class. The 161-amino-acid C-terminal sequence of $EII^{Glc}$ is also similar to that of E. coli enzyme $IIA^{Glc}$, specific for glucose ($EIIA^{Glc}$). The B. ammoniagenes $EII^{Glc}$ consists of three domains; a hydrophobic region (EIIC) and two hydrophilic regions (EIIA, EIIB). The arrangement of structural domains, IIBCA, of the $EII^{Glc}$ is identical to those of EIIs specific for sucrose or ${\beta}$-glucoside. While the domain IIA was removed from the B. ammoniagenes $EII^{Glc}$ the remaining domains IIBC were found to restore the glucose and mannose-utilizing capacity of E. coli mutant lacking $EII^{Glc}$ activity with $EIIA^{Glc}$ of the E. coli mutant. $EII^{Glc}$ contains a histidine residue and a cysteine residue which are putative phosphorylation sites for the protein.

• Korean Journal of Human Ecology
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• v.24 no.2
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• pp.297-325
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• 2015

The purpose of this research is to analyze content elements of Housing area in 2013 middle school Technology Home Economics textbooks 1 and 2, total of 22 according to 2009 revised curriculum. The analysis method was content analysis, focusing on contents of main text, tables & figures, reading material, and activity materials by their content elements. This research will contribute in obtaining suggestion for the post curriculum/textbook development and helping teachers to perform a lesson. The results are as follows. First, Housing area included 1 to 4 content elements per each units, from 2 chapters and 6 units. The content elements stated in Home economics curriculum were total of 16 which consists of 'the meaning of housing,' 'housing types,' 'family forms,' 'family life style cycle,' 'life style,' 'neighboring environment,' 'co-living values,' 'air environment,' 'heat environment,' 'light environment,' 'acoustic environment,' 'space division,' 'circulation', 'effective use of space,' and 'sustainable dwelling practice.' All of these components are dealt with in every textbooks. Second, the numbers of content elements provided in each textbooks were the same, however they showed difference in their contextual aspect. Some contents need supplemental material for their lacking content element. Others need proper understanding of the concept because some showed different contents from the appropriate content elements. Third, repetitions in content elements were observed, the contents of 'co-living values' in textbook 1 and 'sustainable dwelling practice' in textbook 2 were similar in terms of eco-friendly housing, co-housing and universal housing. These two content elements should be merged as one next curriculum, or should be stated together in one subunit.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.4
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• pp.489-498
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• 2018

Objective: Foot and mouth disease (FMD) and porcine reproductive and respiratory syndrome (PRRS) are major diseases that interrupt porcine production. Because they are viral diseases, vaccinations are of only limited effectiveness in preventing outbreaks. To establish an alternative multi-resistant strategy against FMD virus (FMDV) and PRRS virus (PRRSV), the present study introduced two genetic modification techniques to porcine cells. Methods: First, cluster of differentiation 163 (CD163), the PRRSV viral receptor, was edited with the clustered regularly interspaced short palindromic repeats-CRISPR-associated protein 9 technique. The CD163 gene sequences of edited cells and control cells differed. Second, short hairpin RNA (shRNAs) were integrated into the cells. The shRNAs, targeting the 3D gene of FMDV and the open reading frame 7 (ORF7) gene of PRRSV, were transferred into fibroblasts. We also developed an in vitro shRNA verification method with a target gene expression vector. Results: shRNA activity was confirmed in vitro with vectors that expressed the 3D and ORF7 genes in the cells. Cells containing shRNAs showed lower transcript levels than cells with only the expression vectors. The shRNAs were integrated into CD163-edited cells to combine the two techniques, and the viral genes were suppressed in these cells. Conclusion: We established a multi-resistant strategy against viral diseases and an in vitro shRNA verification method.

• Journal of Microbiology and Biotechnology
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• v.14 no.2
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• pp.417-421
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• 2004

A normal prion protein (PrPc) is converted to a protease resistant isoform (PrPsc) by an apparent self-propagating activity in bovine spongiform encephalopathies (BSE), which is a neurodegenerative disease. The cDNA encoding bovine PrP open reading frame (ORP) in Korean cattle was cloned by polymerase chain reaction (PCR). The cloned cDNA had a length of 795 base pairs which coded for a protein of 264 amino acid residues with a calculated molecular mass of 28.6 kDa. Identities of 90, 90, 79 and 78% on nucleotide and 94, 94, 84, and 84% on amino acid sequence were shown to PrP genes from sheep, goat, human, and mouse, respectively. The cloned DNA was ligated into the pQE30 expression vector and transformed into E. coli M15. The PrP was expressed by induction with isopropyl-$\beta$-D-thiogalactoside (IPTG) and purified on the Ni-NTA affinity column. High specific activities of the recombinant PrP were observed in the fraction of pH 5.8 eluate and showed a molecular mass of-29 kDa on SDS-PAGE and Western blot analysis.

• Journal of Microbiology and Biotechnology
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• v.23 no.6
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• pp.759-765
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• 2013

The gene encoding squalene synthase (SQS) of the lipid-producing heterotrophic microalga Aurantiochytrium sp. KRS101 was cloned and characterized. The krsSQS gene is 1,551 bp in length and has two exons and one intron. The open reading frame of the gene is 1,164 bp in length, yielding a polypeptide of 387 predicted amino acid residues with a molecular mass of 42.7 kDa. The deduced krsSQS sequence shares at least four conserved regions known to be required for SQS enzymatic activity in other species. The protein, tagged with $His_6$, was expressed into soluble form in Escherichia coli. The purified protein catalyzed the conversion of farnesyl diphosphate to squalene in the presence of NADPH and $Mg^{2+}$. This is the first report on the characterization of an SQS from a Thraustochytrid microalga.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.6
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• pp.984-991
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• 1997

New tumor suppressor gene, snm23, homologous to human nm23/NDP kinase (human nucleoside diphosphate kinase) gene whose product has a tumor metastasis inhibitory activity, was first cloned from Korean tiger shark (Scyliorhinus forazame) skin cDNA library constructed by using a $\lambda$ ZAP-II cDNA synthesis kit. About $1\times10^5$ plaques were screened and several positive plaques were isolated and confirmed by second screening. The phagemid containing a positive clone from the Uni-Zap XR vector was excised in vivo and the gene containing the tumor metastasis suppressor protein was named as snm23. Cloned gene, snm23, was sequenced with ABI-PRISM 310 Genetic Analyzer. The nucleotide and deduced amino acid sequences of snm23 have shown an open reading frame consisting of 450 base pairs that correspond to a protein of 150 amino acid residues, with a calculated molecular mass of 16.8 kDa. Sequence comparison of snm23 with human nm23/NDP kinase was performed by using Blast protein data base of National Center for Biotechnology Information. In order to determine tissue specificity, reverse transcription-polymerase chain reaction (RT-PCR) was used. Good expression level of snm23/NDP kinase was detected at the tissues from skin, cartilage, and liver of Korean tiger shark.

• The Journal of Korean Medical History
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• v.33 no.2
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• pp.77-87
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• 2020

Objective : The Annals of the Joseon Dynasty is a primary historical record that has provided a great deal of information about what the Joseon Dynasty was like. However, as of yet, we know very little about the medical officers in Joseon dynasty, such as their government posts and official ranks. The purpose of this study is look in to the activities, government posts, and official ranks of the medical personnel by examining Yeongjosillok. Methods : First, I selected historical records containing '醫' in Yeongjosillok. Then, I organized medical officers' name by reading each record. I screened historical records in Yeongjosillok with their names to analyze their activities, government posts, and official ranks. When there was limited information available, I referred to The Daily Records of Royal Secretariat of Joseon Dynasty. Results : I found 262 historical records in Yeongjosillok containing '醫'. Then I found 26 people who served as medical officers in Yeongjosillok. Also, l found that 11 government posts and 7 official ranks were awarded to them throughout the 110 historical records in Yeongjosillok and The Daily Records of Royal Secretariat of Joseon Dynasty. Conclusion : Through this study, I was able to examine the detailed activities of unknown medical officers by studying the historical records in Yeongjosillok and The Daily Records of Royal Secretariat of Joseon Dynasty. Under the Joseon Dynasty's class-based society, the middle class had various restrictions. However, I found that medical officers that belonged to the middle class received exceptional treatment despite their social status.

• Journal of Microbiology and Biotechnology
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• v.26 no.2
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• pp.248-254
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• 2016

A soil metagenome contains the genomes of all microbes included in a soil sample, including those that cannot be cultured. In this study, soil metagenome libraries were searched for microbial genes exhibiting lipolytic activity and those involved in potential lipid metabolism that could yield valuable products in microorganisms. One of the subclones derived from the original fosmid clone, pELP120, was selected for further analysis. A subclone spanning a 3.3 kb DNA fragment was found to encode for lipase/esterase and contained an additional partial open reading frame encoding a wax ester synthase (WES) motif. Consequently, both pELP120 and the full length of the gene potentially encoding WES were sequenced. To determine if the wes gene encoded a functioning WES protein that produced wax esters, gas chromatography-mass spectroscopy was conducted using ethyl acetate extract from an Escherichia coli strain that expressed the wes gene and was grown with hexadecanol. The ethyl acetate extract from this E. coli strain did indeed produce wax ester compounds of various carbon-chain lengths. DNA sequence analysis of the full-length gene revealed that the gene cluster may be derived from a member of Proteobacteria, whereas the clone does not contain any clear phylogenetic markers. These results suggest that the wes gene discovered in this study encodes a functional protein in E. coli and produces wax esters through a heterologous expression system.

• Journal of Microbiology and Biotechnology
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• v.20 no.12
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• pp.1653-1663
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• 2010

The first gene (${\alpha}$-gal1) encoding an extracellular ${\alpha}$-Dgalactosidase from the thermophilic fungus Talaromyces emersonii was cloned and characterized. The ${\alpha}$-gal1 gene consisted of an open reading frame of 1,792 base pairs interrupted by six introns that encoded a mature protein of 452 amino acids, including a 24 amino acid secretory signal sequence. The translated protein had highest identity with other fungal ${\alpha}$-galactosidases belonging to glycosyl hydrolase family 27. The ${\alpha}$-gal1 gene was overexpressed as a secretory protein with an N-terminal histidine tag in the methylotrophic yeast Pichia pastoris. Recombinant ${\alpha}$-Gal1 was secreted into the culture medium as a monomeric glycoprotein with a maximal yield of 10.75 mg/l and purified to homogeneity using Hisbinding nickel-agarose affinity chromatography. The purified enzyme was maximally active at $70^{\circ}C$, pH 4.5, and lost no activity over 10 days at $50^{\circ}C$. ${\alpha}$-Gal1 followed Michaelis-Menten kinetics ($V_{max}\;of\;240.3{\mu}M/min/mg,\;K_m\;of\;0.294 mM$) and was inhibited competitively by galactose ($K_m{^{obs}}$ of 0.57 mM, $K_i$ of 2.77 mM). The recombinant T. emersonii ${\alpha}$-galactosidase displayed broad substrate preference, being active on both oligo- and polymeric substrates, yet had strict specificity for the ${\alpha}$-galactosidic linkage. Owing to its substrate preference and noteworthy stability, ${\alpha}$-Gal1 is of particular interest for possible biotechnological applications involving the processing of plant materials.

• Journal of the East Asian Society of Dietary Life
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• v.13 no.4
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• pp.284-292
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• 2003

This study was conducted to investigate the changes of dietary attitudes and behaviors in relation to the use of computers of elementary school children in Seoul. The total of 451 elementary school children, consisting of 235 females and 216 males, participated in the study. The result of domestic characteristics, dietary attitudes and behaviors, the level of the use of computers, and health-related symptoms of the subjects were achieved through the questionnaires as follows: The average height, weight, BMI and obesity-index of the participants were 149.0 cm, 42.4 kg, 19.0, -8.6, respectively. Anions subjects, 42.8％ answered their bed times were between 11~12 pm, and 82.4％ answered that they had extracurricular activities. The most desired activity as their leisure was computer works (female: 44.3％, male: 62.5％). 38.4％ of children used the computers for 1~2 hours a day and the most general usage of computers was a computer game (66.1％). The changes in dietary habits of the subjects were such as eating faster(30.2％), having lots of snacks(28.8％), eating anything at hand(26.4％), skipping breakfast due to over-sleeping(18.4％). As changes in life patterns, those in the time managements for watching T.V.(35.3％), reading(35.0％), exercising(31.9％), sleeping(27.5％), relaxing(27.5％) and other hobbies(26.4％) were observed. In conclusion, many children were being affected by the socioeconomic factors changing the environments, especially by the need for the use of computers. The rates of eating alone and skipping breakfast were getting higher in the dietary patterns of elementary school children. We found that the changes in social environments according to the heavy use of the computer were affecting on their dietary pattern. The direction and method of nutrition education had to be established for the proper understanding of the desirable dietary behaviors.