• Journal of Korean Elementary Science Education
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• v.20 no.1
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• pp.45-57
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• 2001

The purpose of this study is to guide and apply gettable and accessible materials around everyday life into classroom instruction, and to find out their effects on student academic achievement. In order to achieve it, this study was tried to develop and guide available materials around life in teaming activities, to provide a preliminary assignment that could apply the available materials, and to compare its academic achievement to one from field trip loaming. For the hypothesis testing, first, the experimental and comparing classes were selected based on the results of basic learning diagnostic evaluation, which conducted by the researcher being served as a full-time science teacher of the third and sixth grade of K elementary school in Kwangju for 2 years. Secondly, the instructional use of the available materials around life was applied to the experimental class. Also, the instructional use of the existing materials was applied to the comparing class. Finally, for the testing of the effects on academic achievement, the posterio test was implemented after conducting the experimental instruction in knowledge·understanding, inquiry process, and attitude domains classified by the unit characteristics. Using SPSS/WIN program the t-test was performed in order to compare the differences between the two groups. Major findings were as follows: 1) In the achievement test of knowledge·understanding domain, there were no significant differences at the 5％ level between the experimental and comparing classes. It could be difficult to say, therefore, that the instructional use of the available materials around life was more effective than the instructional use of the existing materials. However, it had some effects on differences between the third grade final achievement test and sixth grade experimental class. 2) In the achievement test of inquiry process domain, there were some significant differences in that the sixth grade experimental class was higher than the third grade out-of classroom experience unit. It was indicated that the instructional use of the available materials around life had some effects on improving the students' inquiry ability 3) In achievement test of attitude domain, there were some significant differences. It was shown that the self-evaluation test of the sixth grade experimental class unit was higher than that of the third grade out-of classroom experience unit; Especially, the learning activities in the experimental class were more active, and the experimental practice ability was improved. It was presented that the instructional use of the available materials around life had some effects on the students' academic achievement in attitude domain. It was concluded from this study that the instructional use of the available materials around life was less effective on Knowledge understanding domain, but was effective on improving their scientific inquiry ability and interest on science education.

• Proceedings of the Korea Society of Environmental Biology Conference
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• 2001.12a
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• pp.89-89
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• 2001

• BMB Reports
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• v.52 no.12
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• pp.700-705
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• 2019

The bacterial effector protein RavZ is secreted by the intracellular pathogen Legionella pneumophila and inhibits host autophagy through an irreversible deconjugation of mammalian ATG8 (mATG8) proteins from autophagosome membranes. However, the roles of the LC3 interacting region (LIR) motifs in RavZ function remain unclear. In this study, we show that a membrane-targeting (MT) domain or the LIR motifs of RavZ play major or minor roles in RavZ function. A RavZ mutant that does not bind to mATG8 delipidated all forms of mATG8-phosphatidylethanolamine (PE) as efficiently as did wild-type RavZ. However, a RavZ mutant with a deletion of the MT domain selectively delipidated mATG8-PE less efficiently than did wild-type RavZ. Taken together, our results suggest that the effects of LIR motifs and the MT domain on RavZ activity are complementary and work through independent pathways.

• Journal of Plant Biotechnology
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• v.32 no.4
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• pp.269-273
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• 2005

Toll-like receptors (TLR) are important components of innate immunity in the defense against pathogens. TLRs recognize pathogen-associated common molecular patterns. TLRs are similar to the receptors involved in defense responses in plants. TLR protein is a type 1 membrane protein, consisting of an extracellular domain containing leucine-rich repeats and a cytoplasmic domain. The cytoplasmic domain delivers ligand recognition signals that result in production of anti-microbial agents. The cytoplasmic domain (amino acid 858-1032) of toll-like receptor 9 has been expressed using methylotrophic yeast Pichia pastoris. The protein expression was confirmed by Western-blot, N-terminal sequencing and MALDl-TOF mass spectrometry. The proteins have been purified by nickel affinity, cation exchange and gel-filtration chromatography.

• Animal Bioscience
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• v.36 no.8
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• pp.1285-1292
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• 2023

Objective: The objective of this study was to develop a novel endolysin (PanLys.1) for the specific killing of the ruminal hyper-ammonia-producing bacterium Peptostreptococcus anaerobius (P. anaerobius). Methods: Whole genome sequences of P. anaerobius strains and related bacteriophages were collected from the National Center for Biotechnology Information database, and the candidate gene for PanLys.1 was isolated based on amino acid sequences and conserved domain database (CDD) analysis. The gene was overexpressed using a pET system in Escherichia coli BL21 (DE3). The lytic activity of PanLys.1 was evaluated under various conditions (dosage, pH, temperature, NaCl, and metal ions) to determine the optimal lytic activity conditions. Finally, the killing activity of PanLys.1 against P. anaerobius was confirmed using an in vitro rumen fermentation system. Results: CDD analysis showed that PanLys.1 has a modular design with a catalytic domain, amidase-2, at the N-terminal, and a cell wall binding domain, from the CW-7 superfamily, at the C-terminal. The lytic activity of PanLys.1 against P. anaerobius was the highest at pH 8.0 (p<0.05) and was maintained at 37℃ to 45℃, and 0 to 250 mM NaCl. The activity of PanLys.1 significantly decreased (p<0.05) after Mn²⁺ or Zn²⁺ treatment. The relative abundance of P. anaerobius did not decrease after administration PanLys.1 under in vitro rumen conditions. Conclusion: The application of PanLys.1 to modulate P. anaerobius in the rumen might not be feasible because its lytic activity was not observed in in vitro rumen system.

• Journal of The Korean Association For Science Education
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• v.32 no.6
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• pp.1039-1049
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• 2012

The first category of Affective Domain objectives in Bloom's Taxonomy is about "Receiving". In it, the first subdivision listed is "Awareness" (Krathwohl, Bloom & Masia, 1964). Since these categories are intended to be hierarchical in ascending order of internalization, it is important that young learners be given ample opportunities in their learning experiences in class to be aware of positive values and effective life skills. This paper reports a feasibility study on the adoption of an integrative cognitive-affective learning approach in a primary school science lesson. 37 primary six students in a Singapore primary school were taught the concept of centre of gravity, including a hands-on activity to find the centre of gravity of an irregularly shaped cardboard by using a plumbline. After reviewing how a plumbline works, their teacher then led them into a discussion on the question "Who is the plumbline in your life?" a reference to identifying positive role models in their lives. From the transcript of the students' in-class sharing and their written responses to the question, it is clear that the integrative cognitive-affective learning approach did enable students to present their ideas and learning experiences in the affective domain quite readily. This conclusion provides a valuable lead to a follow-up project on whether students who are exposed to such integrative learning approaches will be more capable and more aware of identifying important positive social habits or values. If so, then the teaching of values in schools could take on a whole new dimension, that of borrowing students' learning energy in the cognitive domain to learn values and life skills in the affective domain.

• BMB Reports
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• v.54 no.2
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• pp.118-123
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• 2021

The bacterial effector protein RavZ from a pathogen can impair autophagy in the host by delipidating the mammalian autophagy-related gene 8 (mATG8)-phosphatidylethanolamine (PE) on autophagic membranes. In RavZ, the membrane-targeting (MT) domain is an essential function. However, the molecular mechanism of this domain in regulating the intracellular localization of RavZ in cells is unclear. In this study, we found that the fusion of the green fluorescent protein (GFP) to the MT domain of RavZ (GFP-MT) resulted in localization primarily to the cytosol and nucleus, whereas the GFP-fused duplicated-MT domain (GFP-2xMT) localized to Rab5- or Rab7-positive endosomes. Similarly, GFP fusion to the catalytic domain (CA) of RavZ (GFP-CA) resulted in localization primarily to the cytosol and nucleus, even in autophagy-induced cells. However, by adding the MT domain to GFP-CA (GFP-CA-MT), the cooperation of MT and CA led to localization on the Rab5-positive endosomal membranes in a wortmannin-sensitive manner under nutrient-rich conditions, and to autophagic membranes in autophagy-induced cells. In autophagic membranes, GFP-CA-MT delipidated overexpressed or endogenous mATG8-PE. Furthermore, GFP-CA△α3-MT, an α3 helix deletion within the CA domain, failed to localize to the endosomal or autophagic membranes and could not delipidate overexpressed mATG8-PE. Thus, the CA or MT domain alone is insufficient for stable membrane localization in cells, but the cooperation of MT and CA leads to localization to the endosomal and autophagic membranes. In autophagic membranes, the CA domain can delipidate mATG8-PE without requiring substrate recognition mediated by LC3-interacting region (LIR) motifs.

• Proceedings of the Korean Society of Crop Science Conference
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• 2021.10a
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• pp.228-228
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• 2021

• Microbiology and Biotechnology Letters
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• v.34 no.3
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• pp.278-287
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• 2006

Cycloinulooligosaccharide fructanotransferase (CFTase) converts inulin into cycloinulooligosaccharides (cyclofructan, CF) of ${\beta}-(2{\to}1)$-linked D-fructofuranose as well as hydrolysis of cyclofructan. Sequences analysis indicated that CFTase was divided into five distinct regions containing three repeated sequences (R1, R3, and R4) at the N-terminus and C-terminus. Each domain function was investigated by comparison of wild type CFTase enzyme (CFT148) and deletion mutant proteins (CFT108: R1 and R3 deletion; CFT130: R4 deletion; and CFT88: R1, R3, and R4 deletion) of CFTase. The CFT108 mutant had both CFTase and CF hydrolyzing activity as CFT148 did. CFTase activities and CF hydrolysing activities were disappeared in CFT130 and CFT88 mutants. These results indicated that the C-terminal R4 region of P. polymyxa CFTase is necessary for cyclization and hydrolyzing activity.

• BMB Reports
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• v.37 no.6
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• pp.720-725
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• 2004

A number of signaling molecules contain small pleckstrin homology (PH) domains capable of binding phosphoinositides or proteins. Phospholipase C (PLC)-${\gamma}1$ has two putative PH domains, an $NH_2$-terminal (PH1) and a split PH domain ($nPH_2$ and $cPH_2$). We previously reported that the split PH domain of PLC-${\gamma}1$ binds to phosphatidylinositol 4-phosphate (PI(4)P) and phosphatidylinositol 4,5-bisphosphate (PI(4,5)$P_2$) (Chang et al., 2002). To identify the amino acid residues responsible for binding with PI(4)P and PI(4,5)$P_2$, we used site-directed mutagenesis to replace each amino acid in the variable loop-1 (VL-1) region of the PLC-${\gamma}1$ $nPH_2$ domain with alanine (a neutral amino acid). The phosphoinositide-binding affinity of these mutant molecules was analyzed by Dot-blot assay followed by ECL detection. We found that two PLC-${\gamma}1$ nPH2 domain mutants, P500A and H503A, showed reduced affinities for phosphoinositide binding. Furthermore, these mutant PLC-${\gamma}1$ molecules showed reduced PI(4,5)$P_2$ hydrolysis. Using green fluorescent protein (GFP) fusion protein system, we showed that both $PH_1$ and $nPH_2$ domains are responsible for membrane-targeted translocation of PLC-${\gamma}1$ upon serum stimulation. Together, our data reveal that the amino acid residues $Pro^{500}$ and $His^{503}$ are critical for binding of PLC-${\gamma}1$ to one of its substrates, PI(4,5)$P_2$ in the membrane.