• BMB Reports
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• 제44권5호
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• pp.323-328
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• 2011

We screened four B. thuringiensis strains whose parasporal inclusions contained the S-layer protein (SLP), and cloned two slp genes from each strain. Phylogenetic analysis indicated these SLPs could be divided into two groups, SLP1s and SLP2s. To confirm whether SLPs were present in the S-layer or as a parasporal inclusion, strains CTC and BMB1152 were chosen for further study. Western blots with isolated S-layer proteins from strains CTC and BMB1152 in the vegetative phase showed that SLP1s and SLP2s were constituents of the S-layer. Immunofluorescence utilizing spore-inclusion mixtures of strains CTC and BMB1152 in the sporulation phase showed that SLP1s and SLP2s were also constituents of parasporal inclusions. When heterogeneously expressed in the crystal negative strain BMB171, four SLPs from strains CTC and BMB1152 could also form parasporal inclusions. This temporal and spatial expression is not an occasional phenomenon but ubiquitous in B. thuringiensis strains.

• BMB Reports
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• 제44권11호
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• pp.719-724
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• 2011

The $Sec61{\alpha}$ subunit is the core subunit of the protein conducting channel which is required for protein translocation in eukaryotes and prokaryotes. In this study, we cloned a $Sec61{\alpha}$ subunit from Penicillium ochrochloron ($PoSec61{\alpha}$). Sequence and 3D structural model analysis showed that $PoSec61{\alpha}$ conserved the typical characteristics of eukaryotic and prokaryotic $Sec61{\alpha}$ subunit homologues. The pore ring known as the constriction point of the channel is formed by seven hydrophobic amino acids. Two methionine residues from transmembrane ${\alpha}$-helice 7 (TM7) contribute to the pore ring formation and projected notably to the pore area and narrowed the pore compared with the superposed residues at the corresponding positions in the crystal structures or the 3D models of the $Sec61{\alpha}$ subunit homologues in archaea or other eukaryotes, respectively. Results reported herein indicate that the pore ring residues differ among $Sec61{\alpha}$ subunit homologues and two hydrophobic residues in the TM7 contribute to the pore ring formation.

• BMB Reports
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• 제40권1호
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• pp.22-28
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• 2007

MyoD, expressed in skeletal muscle lineages of vertebrate embryo, is one of muscle-specific basic helix-loop-helix (bHLH) transcription factors, which plays a key role in the determination and differentiation of all skeletal muscle lineages. In this study, a cDNA of grass carp MyoD was cloned and characterized from total RNA of grass carp embryos by RT-PCR. The full-length cDNA of grass carp MyoD is 1597 bp. The cDNA sequence analysis reveals an open reading frame of 825 bp coding for a protein of 275 amino acids, which includes a bHLH domain composed of basic domain (1-84th amino acids) and HLH domain (98-142th amino acids), without signal peptide. Then the MyoD cDNA of grass carp was cloned to yeast expression vector pPICZ$\alpha$A and transformed into P. pastoris GS115 strain, the recombinant MyoD protein with a molecular weight of about 31KD was obtained after inducing for 2d with 0.5% methanol in pH 8.0 BMGY medium, and the maximum yield was about 250 mg/L in shaking-flask fermentation. The results were expected to benefit for further studies on the crystal structure and physiological function of fish MyoD.

• 한국전기전자재료학회논문지
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• 제37권2호
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• pp.208-214
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• 2024

This reports the electrical properties of single-crystal β-gallium oxide (β-Ga₂O₃) vertical Schottky barrier diodes (SBDs) with a different guard ring structure. The vertical Schottky barrier diodes (V-SBDs) were fabricated with two types guard ring structures, one is with metal deposited on the Al₂O₃ passivation layer (film guard ring: FGR) and the other is with vias formed in the Al₂O₃ passivation layer to allow the metal to contact the Ga₂O₃ surface (metal guard ring: MGR). The forward current values of FGR and MGR V-SBD are 955 mA and 666 mA at 9 V, respectively, and the specific on-resistance (R_on,sp) is 5.9 mΩ·cm² and 29 mΩ·cm². The series resistance (R_s) in the nonlinear section extracted using Cheung's formula was 6 Ω, 4.8 Ω for FGR V-SBD, 10.7 Ω, 6.7 Ω for MGR V-SBD, respectively, and the breakdown voltage was 528 V for FGR V-SBD and 358 V for MGR V-SBD. Degradation of electrical characteristics of the MGR V-SBD can be attributed to the increased reverse leakage current caused by the guard ring structure, and it is expected that the electrical performance can be improved by preventing premature leakage current when an appropriate reverse voltage is applied to the guard ring area. On the other hand, FGR V-SBD shows overall better electrical properties than MGR V-SBD because Al₂O₃ was widely deposited on the Ga₂O₃ surface, which prevent leakage current on the Ga₂O₃ surface.

• 암석학회지
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• 제6권1호
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• pp.19-33
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• 1997

본역의 지질은 백악기 경상누층군의 퇴적암, 이를 관입 또는 분출한 중성화산암류, 산성화산암류 및 제 4기 충적층으로 구성된다. 중성화산암류는 화성쇄설화산각력암, 석질화산력응회암, 안산암용암으로 구성되며, 산성화산암류는 데사이트질용결응회암 및 유문암용암, 유문암질응회암으로 구성된다. 본 연구에서는 중성 및 산성 화산암류에 대해 암석기재학적 연구와 신선한 시료 10개에 대한 암석화학적 특성을 고찰하고, K-Ar법에 의한 절대연대 측정을 실시하였다. 현미경 관찰에서 안산암용암은 사장석이 주 반정광물로 나타나며, 기질은 미정질 내지 은미정질로서 반정광물과 동일한 필로택시틱 조직을 보인다. 안산암질각력암은 퇴적암 및 안산암의 자력 암편을 포함한다. 테사이트질용결응회암은 현저한 파라택시틱 조직을 보이며, 유문암용암은 유상구조를 잘 보여 주고, 안산암의 암편을 함유하고 용결구조가 현저한 화산력용결응회암이 나타나는데, 유문암용암에서 기질의 함량은 80.9~ 89.3%에 이른다. 주 반정광물은 사장석이며 부분적으로 녹염석, 녹니석, 방해석, 제오라이트, 푸로필라이트 등으로 2차 변질되어 나타난다. 본역의 화산암류는 Norm값에 의한 Q-A-P 도표에서 현무암질안산암, 안산암, 데사이트, 유문암의 일련의 분화과정을 나타내고 대부눈 칼크알칼리 계열에 속한다. 화산암류의 화학조성은 $SiO_2$ 함량이 57.61~75.40 %이며, MgO, CaO, $Fe_2O_3$, $Al_2O_3$, $Ti_2$, MnO, $P_2O_5$ 등은 $SiO_2$함량이 증가함에 따라 연속적으로 증가하는 경향을 보인다. 주성분원소 및 미량 원소의 변화도에서 안산암질에서 유문암으로 분화되는, 즉 마그마의 정출 분화 특징을 뚜렷이 보여준다. REE 양상 및 spider 도표에서 일정한 분화 경향을 보이며 나란하다. spider 도표에서 본역의 화산암류는 Th, La, Nd, Gd 등이 부화되어 있으며, Ba, Nb, Sr, Hf, Zr 등이 결핍되어 있는 특징을 나타낸다. 안산암에서 유문암으로 분화가 진행될수록 Cs, Sr, Eu이 점차 결핍되는 경향이, Th가 증가하는 경향이 있고, Ba, Nb, Sr, Eu의 부(-)의 이상값이 점차 커지는 경향을 보인다. 희유 원소의 변화 경향에서 안산암과 중성 암맥, 데사이트와 유문암의 경향이 서로 일치함을 볼 수 있다. 주성분 원소 및 미량 원소 함량 변화는 본역의 화산암이 안산암으로부터 일연의 분별결정작용 산물임을 암시하며, 또한 $Al_2O_3$와 CaO 함량의 관계도, Th/Yb 비에 대한 Ta/Yb 비의 관계도, $Ce_N/Yb_N$ 대 $Ce_N$의 관계도에 따른 판별에서도 분별결정작용의 경향을 따르고 있다. 본역의 화산암은 $K_2O$, $Na_2O$, CaO 삼각도에서 도호의 영역에, Ba/La비, La/Th비에 의한 판별도에서 조산대의 high-K suite에 속한다. Rb 대 (Y+Nb)의 판별도 및 Hf-Th-Ta 지구조 판별도에서 지판이 침강 섭입하는 지판 경계부(destructive plate margin) 중 화산호의 조구적 영역에 도시된다. 본역의 화산암을 생성시킨 마그마는 $Al_2O_3$와 CaO 함량의 관계도, mode에서 나타나는 사장석 반정, 분화가 진행될수록 부의 Eu 이상이 증가하는 것 등에서 사장석의 분별이 우세한 분별결정작용을 거쳤음을 알 수 있다. 안산암질암을 관입한 중상 암맥에서 측정한 암석 년령은 $97.0{\pm}6.8~94.5{\pm}6.6$, 데사이트질암은 $68.9{\pm}4.8,\61.5{\pm}4.9~60.7{\pm}4.2$Ma으로 측정되었고, 이것은 백악기 유천층군과 대비되며, 백악기 유천층군 암석의 지화학적 자료와 본역 화산암의 지화하적 자료는 판별도 등에서 같은 영역에 도시된다.

• BMB Reports
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• 제40권5호
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• pp.773-782
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• 2007

Different isolates of the soil bacterium Bacillus thuringiensis produce multiple crystal (Cry) proteins toxic to a variety of insects, nematodes and protozoans. These insecticidal Cry toxins are known to be active against specific insect orders, being harmless to mammals, birds, amphibians, and reptiles. Due to these characteristics, genes encoding several Cry toxins have been engineered in order to be expressed by a variety of crop plants to control insectpests. The cotton boll weevil, Anthonomus grandis, and the fall armyworm, Spodoptera frugiperda, are the major economically devastating pests of cotton crop in Brazil, causing severe losses, mainly due to their endophytic habit, which results in damages to the cotton boll and floral bud structures. A cry1Ia-type gene, designated cry1Ia12, was isolated and cloned from the Bt S811 strain. Nucleotide sequencing of the cry1Ia12 gene revealed an open reading frame of 2160 bp, encoding a protein of 719 amino acid residues in length, with a predicted molecular mass of 81 kDa. The amino acid sequence of Cry1Ia12 is 99% identical to the known Cry1Ia proteins and differs from them only in one or two amino acid residues positioned along the three domains involved in the insecticidal activity of the toxin. The recombinant Cry1Ia12 protein, corresponding to the cry1Ia12 gene expressed in Escherichia coli cells, showed moderate toxicity towards first instar larvae of both cotton boll weevil and fall armyworm. The highest concentration of the recombinant Cry1Ia12 tested to achieve the maximum toxicities against cotton boll weevil larvae and fall armyworm larvae were 230 ${\mu}g/mL$ and 5 ${\mu}g/mL$, respectively. The herein demonstrated insecticidal activity of the recombinant Cry1Ia12 toxin against cotton boll weevil and fall armyworm larvae opens promising perspectives for the genetic engineering of cotton crop resistant to both these devastating pests in Brazil.

• BMB Reports
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• 제40권5호
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• pp.625-635
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• 2007

The enzyme squalene synthase (EC 2.5.1.21) catalyzes a reductive dimerization of two farnesyl diphosphate (FPP) molecules into squalene, a key precursor for the sterol and triterpene biosynthesis. A full-length cDNA encoding squalene synthase (designated as TcSqS) was isolated from Taxus cuspidata, a kind of important medicinal plants producing potent anti-cancer drug, taxol. The full-length cDNA of TcSqS was 1765 bp and contained a 1230 bp open reading frame (ORF) encoding a polypeptide of 409 amino acids. Bioinformatic analysis revealed that the deduced TcSqS protein had high similarity with other plant squalene synthases and a predicted crystal structure similar to other class I isoprenoid biosynthetic enzymes. Southern blot analysis revealed that there was one copy of TcSqS gene in the genome of T. cuspidata. Semi-quantitative RT-PCR analysis and northern blotting analysis showed that TcSqS expressed constitutively in all tested tissues, with the highest expression in roots. The promoter region of TcSqS was also isolated by genomic walking and analysis showed that several cis-acting elements were present in the promoter region. The results of treatment experiments by different signaling components including methyl-jasmonate, salicylic acid and gibberellin revealed that the TcSqS expression level of treated cells had a prominent diversity to that of control, which was consistent with the prediction results of TcSqS promoter region in the PlantCARE database.

• BMB Reports
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• 제37권3호
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• pp.304-313
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• 2004

Three-dimensional (3D) models for the 65-kDa activated Cry4A and Cry4B $\delta$-endotoxins from Bacillus thuringiensis subsp. israelensis that are specifically toxic to mosquito-larvae were constructed by homology modeling, based on atomic coordinates of the Cry1Aa and Cry3Aa crystal structures. They were structurally similar to the known structures, both derived 3D models displayed a three-domain organization: the N-terminal domain (I) is a seven-helix bundle, while the middle and C-terminal domains are primarily comprise of anti-parallel $\beta$-sheets. Circular dichroism spectroscopy confirmed the secondary structural contents of the two homology-based Cry4 structures. A structural analysis of both Cry4 models revealed the following: (a) Residues Arg-235 and Arg-203 are located in the interhelical 5/6 loop within the domain I of Cry4A and Cry4B, respectively. Both are solvent exposed. This suggests that they are susceptible to tryptic cleavage. (b) The unique disulphide bond, together with a proline-rich region within the long loop connecting ${\alpha}4$ and ${\alpha}5$ of Cry4A, were identified. This implies their functional significance for membrane insertion. (c) Significant structural differences between both models were found within domain II that may reflect their different activity spectra. Structural insights from this molecular modeling study would therefore increase our understanding of the mechanic aspects of these two closely related mosquito-larvicidal proteins.

• BMB Reports
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• 제40권3호
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• pp.315-324
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• 2007

The novel $\alpha$-amylase purified from locally isolated strain, Bacillus sp. KR-8104, (KRA) (Enzyme Microb Technol; 2005; 36: 666-671) is active in a wide range of pH. The enzyme maximum activity is at pH 4.0 and it retains 90% of activity at pH 3.5. The irreversible thermoinactivation patterns of KRA and the enzyme activity are not changed in the presence and absence of $Ca^{2+}$ and EDTA. Therefore, KRA acts as a $Ca^{2+}$-independent enzyme. Based on circular dichroism (CD) data from thermal unfolding of the enzyme recorded at 222 nm, addition of $Ca^{2+}$ and EDTA similar to its irreversible thermoinactivation, does not influence the thermal denaturation of the enzyme and its Tm. The amino acid sequence of KRA was obtained from the nucleotide sequencing of PCR products of encoding gene. The deduced amino acid sequence of the enzyme revealed a very high sequence homology to Bacillus amyloliquefaciens (BAA) (85% identity, 90% similarity) and Bacillus licheniformis $\alpha$-amylases (BLA) (81% identity, 88% similarity). To elucidate and understand these characteristics of the $\alpha$-amylase, a model of 3D structure of KRA was constructed using the crystal structure of the mutant of BLA as the platform and refined with a molecular dynamics (MD) simulation program. Interestingly enough, there is only one amino acid substitution for KRA in comparison with BLA and BAA in the region involved in the calcium-binding sites. On the other hand, there are many amino acid differences between BLA and KRA at the interface of A and B domains and around the metal triad and active site area. These alterations could have a role in stabilizing the native structure of the loop in the active site cleft and maintenance and stabilization of the putative metal triad-binding site. The amino acid differences at the active site cleft and around the catalytic residues might affect their pKa values and consequently shift its pH profile. In addition, the intrinsic fluorescence intensity of the enzyme at 350 nm does not show considerable change at pH 3.5-7.0.

• Tuberculosis and Respiratory Diseases
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• 제78권4호
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• pp.440-444
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• 2015

Gastric mucosal damage by iron pills is often reported. However, iron pill aspiration is uncommon. Oxidation of the impacted iron pill causes bronchial mucosal damage that progresses to chronic bronchial inflammation, necrosis, endobronchial stenosis and rarely, perforation. We reported a case of a 92-year-old woman with chronic productive cough and significant left-sided atelectasis. Bronchoscopy revealed substantial luminal narrowing with exudative inflammation of the left main bronchus. Bronchial washing cytology showed necroinflammatory exudate and a small amount of brown material. Mucosal biopsy showed diffuse brown pigments indicative of ferrous pigments, crystal deposition, and marked tissue degeneration. After vigorous coughing, she expectorated dark sediments and her symptoms and radiological abnormalities improved. There are a few such reports worldwide; however, this was the first case reported in Korea. Careful observation of aspiration-prone patients and early detection of iron pill aspiration may prevent iron pill-induced bronchial injury.