• 환경생물
• /
• 제25권4호
• /
• pp.289-296
• /
• 2007

본 연구에서는 내분비계 장애물질으로 알려져 있고, 플라스틱 제품의 가소제로 사용되고 있는 di-(2-ethylhexyl) phthalate (DEHP)가 흰쥐의 간세포 미세구조와 간조직내 metallothionein (MT)의 발현 양상에 미치는 영향을 조사하였다. DEHP는 흰쥐 간세포의 미세구조와 MT 발현에 영향을 주었다. 실험군의 경우 조면소포체가 발달하고, 미토콘드리아가 증가하며 리소솜 혹은 퍼옥시좀들이 증가하는 경향을 나타냈다. 한편, MT 발현의 변화를 면역세포 화학적 방법과 western Blot을 수행한 결과 저농도 투여군에서는 약 1.5배, 고농도 투여군에서는 약 2배 가량 증가하는 결과를 나타냈다. 결론적으로 DEHP는 흰쥐 간세포의 구조와 기능에 영향을 주는 것으로 판단되며, 이러한 세포내 스트레스가 MT 발현 증가 현상과 연관이 있을 것으로 판단된다.

• BMB Reports
• /
• 제40권2호
• /
• pp.239-246
• /
• 2007

Riboflavin synthase from Escherichia coli is a homotrimer of 23.4 kDa subunits and catalyzes the formation of one molecule each of riboflavin and 5-amino-6-ribitylamino- 2,4(1H,3H)-pyrimidinedione by the transfer of a 4-carbon moiety between two molecules of the substrate, 6,7- dimethyl-8-ribityllumazine. Each subunit comprises two closely similar folding domains. Recombinant expression of the N-terminal domain is known to provide a $C_2$-symmetric homodimer. In this study, the binding properties of wild type as well as two mutated proteins of N-terminal domain of riboflavin synthase with various ligands were tested. The replacement of the amino acid residue A43, located in the second shell of riboflavin synthase active center, in the recombinant N-terminal domain dimer reduces the affinity for 6,7-dimethyl-8-ribityllumazine. The mutation of the amino acid residue C48 forming part of activity cavity of the enzyme causes significant $^{19}F$ NMR chemical shift modulation of trifluoromethyl derivatives of 6,7-dimethyl-8-ribityllumazine in complex with the protein, while substitution of A43 results in smaller chemical shift changes.

• International Journal of Industrial Entomology and Biomaterials
• /
• 제13권1호
• /
• pp.23-30
• /
• 2006

BmCu/Zn SOD was isolated from early embryo of Bombyx mori using microarray analysis. The BmCu/Zn SOD gene was observed during the early embryonic stage with the strongest signal found at the unfertilizaion, fertilization and blastoderm stages. The BmCu/Zn SOD gene encodes a protein of 154 amino acids with a calculated Mr of 15 kDa. The deduced amino acid sequence of BmCu/Zn SOD indicated that the residues that form on the Cu/Zn binding site are conserved and that the sequence is a 60% identity to that of M. domestica. In a phylogenetic tree, Bm SOD was also close to Drosophila SODs rather than other insect SODs. The BmCu/Zn SOD gene exists as a single copy in the genome. Transcripts of BmCu/Zn SOD cDNA were identified by northern blot analysis. The expression of the BmCu/Zn SOD gene was observed weakly in most of larvae, pre-pupae, pupae and adult tissues. Also, the BmCu/Zn SOD gene was observed in early embryonic stage. Although the roles of SODs remains to be further elucidated, the high expression of BmCu/Zn SOD gene at before 24 h post fertilization suggests that this gene is of general importance during early embryogenesis in the Bombyx mod.

• Journal of Microbiology and Biotechnology
• /
• 제8권6호
• /
• pp.676-684
• /
• 1998

Baculovirus transfer and expression vectors with Hyphantria cunea nuclear polyhedrosis virus (HcNPV) were constructed. An initial transfer vector, pHcEV, constructed using HcNPV was previously reported (Park et al. 1993. J. Kor. Soc. Viral. 23: 141-151). Herein, the size of the vector was properly reduced, and a functionally perfect vector was constructed and named pHcEV-IV (6.7 kb). The vector has a 2.2-kb HcNPV DNA sequence in the 5'-flanking region of the vector's polyhedrin gene promoter. The 1.8-kb HcNPV DNA sequence, poly A signal sequence, T3 primer sequence, and 13 multicloning site sequences, in order, were ligated in front of the translation start codon of the polyhedrin gene. The cloning indicating marker lacZ gene was inserted into the pHcEV-IV, named pHcEV-IV-lacZ, and transferred into the wild-type virus. Recombinant expression virus, lacZ-HcNPV, was constructed by replacing the lacZ gene in the pHcEV-IV-lacZ with the polyhedrin gene of the wild-type virus. The recombinant virus was isolated from blue plaques that produce $\beta$-galactosidase without polyhedra. The lacZ gene insertion was confirmed by Southern hybridization analysis. The expression of the lacZ gene in Spodoptera frugiperda cells infected with the lacZ-HcNPV was examined by SDS-PAGE and colorimetric assay. One 116-kDa LacZ protein band appeared on the PAGE. The production rate of the $\beta$-galactosidase was approximately 50 international units (IU) per min per ml between 2 to 5 days postinfection (p.i.). The highest activity occurred at five days p.i. was 170 IU/min/$m\ell$. The enzyme activity first appeared about 20 h p.i. as measured by colorimetric assay.

• 한국임상수의학회지
• /
• 제27권1호
• /
• pp.23-28
• /
• 2010

Mycobacterium avium ssp paratuberculosis, intracellular bacteria that can cause chronic granulomatous enteritis in cattle, continues to pose significant economic losses and health problem with high prevalence. The purpose of this study is the polymerase chain reaction (PCR)-base strategy for early detection of M. avium ssp paratuberculosis in whole blood. Blood samples were collected from korean cattles in Jeonbuk, Korea. The 16 out of 88 serum samples were detected M. partuberculosis by ELISA. Then samples of infected 8 Korean cattles were amplified by PCR. The PCR amplified targets are 16s rDNA and heat shock protein 65kDa (hsp 65). The 16s rDNA provided a highly sensitive and specific tool for the direct detection of mycobacteria. In addition M. avium was confirmed characteristically by the hsp65. Finally there were sure to M. avium ssp paratuberculosis by IS900 PCR. The restriction fragment length polymorphism was identified by PCR amplifications and subsequence restriction enzyme digestions with Pst I of a hsp65. These results indicate that confirm M. avium with 16s rDNA, hsp65 and a restriction fragment length polymorphism in the hsp65 gene can be seem the other pattern. Therefore, these results can be used for clinical direct detections of M. avium ssp paratuberculosis in whole blood of Korean cattle and also to be used epidemiological researches.

• Clinical and Experimental Pediatrics
• /
• 제58권9호
• /
• pp.341-346
• /
• 2015

Purpose: To evaluate the practical applications of the diagnosis algorithms recommended by the American Academy of Pediatrics urinary tract infection (UTI) guideline. Methods: We retrospectively reviewed the medical records of febrile UTI patients aged between 2 and 24 months. The patients were divided into 3 groups: group I (patients with positive urine culture and urinalysis findings), group II (those with positive urine culture but negative urinalysis findings), and group III (those with negative urine culture but positive urinalysis findings). Clinical, laboratory, and imaging results were analyzed and compared between the groups. Results: A total of 300 children were enrolled. The serum C-reactive protein level was lower in children in group II than in those in groups I and III (P<0.05). Children in group I showed a higher frequency of hydronephrosis than those in groups II and III (P<0.05). However, the frequencies of acute pyelonephritis (APN), vesicoureteral reflux (VUR), renal scar, and UTI recurrence were not different between the groups. In group I, recurrence of UTI and presence of APN were associated with the incidence of VUR (recurrence vs. no recurrence: 40% vs.11.4%; APN vs. no APN: 23.3% vs. 9.2%; P<0.05). The incidence of VUR and APN was not related to the presence of hydronephrosis. Conclusion: UTI in febrile children cannot be ruled out solely on the basis of positive urinalysis or urine culture findings. Recurrence of UTI and presence of APN may be reasonable indicators of the presence of VUR.

• 한국식물학회:학술대회논문집
• /
• 한국식물학회 1999년도 Proceedings of the 17th Symposium on Plant Biology Environmental Stress and Photosynthesis
• /
• pp.83-90
• /
• 1999

Electron crystallography of two-dimensional crystalsand electron cryo-microscopy is becoming an established method for determining the structure and function of a variety of membrane proteins that are providing difficult to crystallize in three dimension. In this study this technique has been used to investigate the structure of a ~160 kDa reaction centre sub-core complex of photosystem II. Photosystem II is a photosynthetic membrane protein consisting of more than 25 subunits. It uses solar energy to split water releasing molecular oxygen into the atmosphere and creates electrochemical potential across the thylakoid membrane, which is eventually utilized to generate ATP and NADPH. Images were taken using Philips CM200 field emission gun electron microscope with an acceleration voltage of 200kW at liquid nitrogen temperature. In total, 79 images recorded dat tilt angles ranging from 0 to 67 degree yielded amplitudes and phases for a three-dimensional map with an in-plant resolution of 6$\AA$ and 11.4$\AA$ in the third dimension shows at least 23 transmembrane helices resolved in a monomeric complex, of which 18 were able to be assigned to the D1, D2, CP47 , and cytochrome b559 alfa beta-subunits with their associated pigments that ae active in electron transport (Rhee, 1998, Ph.D.thesis). The D1/D2 heterodimer is located in the central position within the complex and its helical scalffold is remarkably similar to that of the reaction centres not only in purple bacteria but also in plant photosystem I (PSI) , indicating a common evoluationary origin of all types of reaction centre in photosynthetic organism known today 9RHee et al. 1998). The structural homology is now extended to the inner antenna subunit, ascribed to CP47 in our map, where the 6 transmembrane helices show a striking structural similarity to the corresponding helices of the PSI reaction centre proteins. The overall arrangement of the chlorophylls in the D1 /D2 heterodimer, and in particular the distance between the central pair, is ocnsistent with the weak exciton coupling of P680 that distinguishes this reaction centre from bacterial counterpart. The map in most progress towards high resolution structure will be presented and discussed.

• 한국식생활문화학회지
• /
• 제30권1호
• /
• pp.77-85
• /
• 2015

This study investigated the physicochemical properties and antioxidative activity of soymilk added with buckwheat sprout (SBS), and sensory properties of SBS were analyzed with control soymilk (CS). Moisture content decreased while protein, ash, and lipid contents increased according to content of buckwheat sprout. Solids content and viscosity tended to increase with addition of buckwheat sprout from 0% (CS) to 2.5% (2.5SBS), 3.5% (3.5SBS), and 4.5% (4.5SBS). SBS was found the significant pH drop from 7.08 to 6.43, 6.34, and 6.21. Suspension stability of soymilk slightly decreased with addition of buckwheat sprout. Hunter's color value L of SBS decreased while a and b values increased in comparison with CS. Rutin content was measured 23.78 mg/100 g (2.5SBS), 39.68 mg/100 g (3.5SBS), and 44.80mg/100 g (4.5SBS). Vitamin C content in SBS was higher than CS. Daidzin content increased as buckwheat sprout was added. Total phenolic content increased from 100.95 mg/100 g (CS) up to 315.71 mg/100 g (4.5SBS). For free radical scavenging activity, SBS was significantly higher than CS. In conclusion, SBS can be used as a functional food with higher amounts of rutin, vitamin C, total phenolics, and daidzin. 2.5SBS and 3.5SBS also showed better overall quality characteristics.

• 한국작물학회지
• /
• 제60권3호
• /
• pp.394-400
• /
• 2015

본 연구는 2배체와 4배체 도라지 단백질 발현지도 제작과 도라지의 생리활성에 관여하는 메카니즘을 분자적 수준에서 해석하기 위한 기초자료를 얻고자 실시하였다. 2배체 및 4배체 도라지의 단백질의 발현 양상은 배수성에 따른 특이성은 관찰 할 수 없었으며, 모두 분자량 15~100 kDa 크기, pH 4.0~8.0의 범위에 분포하는 것으로 나타났다. 동정된 39개의 단백질 중 2배체에 비해 4배체에서 2개의 단백질이 up-regulated 되었고, 37개의 단백질이 down-regulated 되었다. 단백질을 기능별로 분류한 결과, 산화환원효소의 활성(oxidoreductase activity)기능을 갖는 단백질이 23.7%의 비율로 가장 많았고 다음은 nucleotide binding 기능의 단백질이 15.8%의 비율로 높았다.

• 한국작물학회지
• /
• 제59권3호
• /
• pp.319-324
• /
• 2014

콩 성숙 종자에 존재하는 raffinose 및 stachyose의 함량이 적은 콩 품종 육성을 위하여 요구되는 기본정보를 얻기 위하여 장려품종, 육성계통, 유전자원등 22개의 유전자원과 2년간 함량변이를 관찰한 결과는 아래와 같았다. 1. Raffinose의 함량(g/kg)은 1년차의 경우 $2.68{\pm}0.21-5.87{\pm}2.43$의 범위를 보였고 2년차에서는 $3.24{\pm}0.37-9.05{\pm}0.16$의 범위를 보였다. 2. Stachyose의 함량(g/kg)은 1년차에서 $4.23{\pm}0.98-27.68{\pm}9.90$, 2년차에서는 $5.11{\pm}1.09-25.32{\pm}0.35$의 범위를 보였다. 3. 분산분석 결과 유전자형과 년차간 stachyose 및 raffinose의 함량은 고도의 유의성을 보였고 상호작용은 인정되지 않았다. 4. Stachyose 함량에서는 2년차 평가 모두에서 Da-7, 116-13, RS-78 유전자형이 유의하게 낮은 함량을 보였다. 5. Raffinose 함량은 1년차 평가에서 Hwangkumkong 및 P34-162 유전자형이 낮은 값을 보였고, 2년차에서는 116-13과 Da-7 유전자형이 낮은 값을 보였다. 6. Raffinose와 stachyose의 함량간에는 2년차 평가에서 유의한 정의 상관관계($R^2=0.1985^*$)을 보였다.