• 대한수의학회지
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• 제43권3호
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• pp.339-347
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• 2003

The regional distribution and relative frequency of some endocrine cells in the pancreas of the Korean aucha perch, Coreoperca herzi Herzenstein belonging to the family Serranidae in order Perciformis, were observed using specific mammalian antisera against serotonin, insulin, glucagon, somatostatin and human pancreatic polypeptide (hPP) by peroxidase antiperoxidase (PAP) method. The pancreas was divided into four portions (principal and secondary islets, exocrine and pancreatic duct regions). In addition, the pancreatic islet regions were further subdivided into three regions (central, mantle and peripheral regions). Spherical to spindle or occasionally round to oval immunoreactive (IR) cells were demonstrated in the pancreatic islets and exoccrine portions, but no cells were detected in the pancreatic duct portions. In the principal islets, serotonin-IR cells were not detected but most of insulin-IR cells were located in the central regions and they were also demonstrated in the mantle and peripheral regions in moderate and rare frequencies, respectively. Glucagon- and hPP-IR cells were mainly situated in the mantle regions but the cells were also demonstrated in the peripheral regions in relatively lower frequency. Somatostatin-IR cells were evenly distributed in the central and mantle regions in a few frequency and cells were also demonstrated in the peripheral regions in rare frequency. Cell clusters were consisted of hPP-IR cells that were situated in the peripheral to mantle regions. In the secondary islet portions, serotonin-IR cells were randomly distributed throughout the whole pancreatic islet regions but lower frequency was detected in the peripheral regions compared to that in central and mantle regions where cells were detected in a few frequency, respectively. Insulin-IR cells were restricted to the central regions in numerous frequency and glucagon-IR cells were evenly distributed in the mantle and peripheral regions in moderate frequencies, respectively. Somatostatin-IR cells were observed in the central and mantle regions in moderate and a few frequencies, respectively. In addition, hPP-IR cells showed similar distributional patterns to those of glucagon-IR cells except cells were also located in the central regions in rare frequency. In the exocrine portions, only glucagon- and hPP-IR cells were demonstrated in rare and a few frequencies, respectively. In conclusion, the regional distribution and relative frequency of pancreatic endocrine cells of the Korean aucha perch showed general patterns, which were observed in other teleost. However, some species-dependent different distributional patterns and/or relative frequencies were also demonstrated especially to serotonin-IR cells. In pancreas of the Korean aucha perch, insulin-IR cells were the most predominant cell type followed by glucagon-, somatostatin-, hPP- and serotonin-IR cells.

• Journal of Microbiology
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• 제44권1호
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• pp.54-63
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• 2006

Ferritin is a major eukaryotic protein and in humans is the protein of iron storage. A partial gene fragment of ferritin (255 bp) taken from the total RNA of Periserrula leucophryna, was amplified by RT-PCR using oligonucleotide primers designed from the conserved metal binding domain of eukaryotic ferritin and confirmed by DNA sequencing. Using the $^{32}P-labeled$ partial ferritin cDNA fragment, 28 different clones were obtained by the screening of the P. leucophryna cDNA library prepared in the Uni-ZAP XR vector, sequenced and characterized. The longest clone was named the PLF (Periserrula leucophryna ferritin) gene and the nucleotide and amino acid sequences of this novel gene were deposited in the GenBank databases with accession numbers DQ207752 and ABA55730, respectively. The entire cDNA of PLF clone was 1109 bp (CDS: 129-653), including a coding nucleotide sequence of 525 bp, a 5' -untranslated region of 128 bp, and a 3'-noncoding region of 456 bp. The 5'-UTR contains a putative iron responsive element (IRE) sequence. Ferritin has an open reading frame encoding a polypeptide of 174 amino acids including a hydrophobic signal peptide of 17 amino acids. The predicted molecular weights of the immature and mature ferritin were calculated to be 20.3 kDa and 18.2 kDa, respectively. The region encoding the mature ferritin was subcloned into the pT7-7 expression vector after PCR amplification using the designed primers and included the initiation and termination codons; the recombinant clones were expressed in E. coli BL21(DE3) or E. coli BL21(DE3)pLysE. SDS-PAGE and western blot analysis showed that a ferritin of approximately 18 kDa (mature form) was produced and that by iron staining in native PAGE, it is likely that the recombinant ferritin is correctly folded and assembled into a homopolymer composed of a single subunit.

• 대한한방내과학회지
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• 제29권2호
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• pp.469-489
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• 2008

Objectives : The purpose of this study was to investigate the effects of Chungganhaeju-tang(Qingganjiejiu-tang) on alcoholic liver damaged by applying proteomics. Materials and Methods : Sprague-Dawley rats were used in this experiment the rats were divided into the normal group, the control group(alcohol) and the sample group(CGHJT +alcohol). The ethanol was orally administered twice a day for 6 weeks in the control and sample groups. Water instead of ethanol was orally administered twice a day for 6 weeks in the normal group. CGHJT extract was orally administered once a day for 6 weeks in the sample group. The livers of each group were processed and assessed by histology, Western Blot, $Oxyblot^{TM}$, CBB and 2-dimensional electrophoresis. Results : In the histological findings of the liver, CGHJT inhibited hepatic fibrogenesis induced by alcohol. TIMP-1 decreased in the sample group assessed by western blot and statistical significance was noted by dot blotting(p<0.05). In the $Oxyblot^{TM}$, protein oxidation induced by alcohol treatment decreased with CGHJT. In the 2-dimensional electrophoresis finding, increased proteins alcohol such as HSP 60, 60kDa heat shock protein, 3-mercaptopyruvate sulfurtransferase were normalized by CGHJT. CGHJT was considered to normalize the anti-oxidation activity elevated by alcohol. In the 2-dimensional electrophoresis finding, increased oxidized proteins such as actin, prolyl 4-hydroxylase beta polypeptide, 94kDa glucose regulated protein(GRP94), heat shock protein 90-alpha(HSC86), calreticulin precursor(CRP55), ATP synthase beta chain mitochondrial precursor, caspase-8 precursor, and dihydrolipoamide succinyltransferase(E2) decreased with CGHJT. CGHJT was considered to reduce the oxidative stress of alcohol. Conclusion : Chungganhaeju-tang(Qingganjiejiu-tang) exerts an inhibitory effect against the fibrosis and protein oxidation induced by alcohol treatment of rat liver. CGHJT was considered to normalize the elevated anti-oxidation activity by alcohol and to reduce the level of oxidative stress due to alcohol.

• BMB Reports
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• 제39권2호
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• pp.158-166
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• 2006

In many organisms, trehalose acts as protective metabolite against harsh environmental stresses, such as freezing, drought, nutrient starvation, heat and salt. Herein a cDNA (designated as GbTPS, GenBank Accession Number AY884150) encoding a trehalose-6-phosphate synthase homologue was isolated and characterized from the living fossil plant, Ginkgo biloba, which is highly tolerant to drought and cold. GbTPS encoded an 868-amino-acid polypeptide with a predicted isoelectric point of 5.83 and molecular mass of 97.9 kD. Amino acid sequence alignment revealed that GbTPS shared high identity with class II trehalose-6-phosphate synthase homologues (67% identical to AtTPS7), but had only 17% and 23% of identity with OstA from Escherichia coli and ScTPS1 from S. cerevisiae, respectively. DNA gel blot analysis indicated that GbTPS belonged to a small multi-gene family. The expression analysis by RT-PCR showed that GbTPS expressed in a tissue-specific manner in G biloba and might involve in leaf development. GbTPS was also found to be induced by a variety of stresses including cold, salt, drought and mannitol.

• BMB Reports
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• 제39권1호
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• pp.68-75
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• 2006

In higher plants, P450s participate in the biosynthesis of many important secondary metabolites. Here we reported for the first time the isolation of a new cytochrome P450 cDNA that expressed in a stem-specific manner from Camptotheca acuminata (designated as CaSS), a native medicinal plant species in China, using RACE-PCR. The full-length cDNA of CaSS was 1735 bp long containing a 1530 bp open reading frame (ORF) encoding a polypeptide of 509 amino acids. Bioinformatic analysis revealed that CASS contained a heme-binding domain PFGXGRRXCX and showed homology to other plant cytochrome P450 monooxygenases and hydroxylases. Southern blotting analysis revealed that there was only one copy of the CaSS present in the genome of Camptotheca acuminata. Northern blotting analysis revealed that CaSS expressed, in a tissue-specific manner, highly in stem and lowly in root, leaf and flower. Our study suggests that CaSS is likely to be involved in the phenylpropanoid pathway.

• 대한토목학회논문집
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• 제14권3호
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• pp.679-688
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• 1994

본 연구에서는 UASB 반응조의 형상변화에 따른 입상슬러지의 물리 화학적 및 형태학적인 특성이 조사되었다. 또한 반응조내의 수소분압의 크기에 따른 반응조운전의 안정성이 조사되었다. 수소분압이 높게 유지된 수정개발된 UASB 반응조의 경우가 상대적으로 수소분압이 낮게 유지된 일반적인 UASB 반응조의 경우에 비해 입상슬러지의 침전성 및 미생물보유능이 더 우수하게 나타났다. 입상슬러지의 형성과 그 안정성에 수소분압이 큰 영향을 미치는 것으로 나타났다. 입상슬러지의 화학적 조성식은 일반적인 UASB 반응조와 수정개발된 UASB 반응조가 각각 $C_7H_{12}O_{4.6}N$과 $C_5H_9O_3N$으로, 일반적인 미생물의 경험식인 $C_5H_7O_2N$과는 상이하게 나타났다. 특히 수정개발된 UASB 반응조의 경우 입상슬러지내에 질소성분이 일반적인 혐기성 미생물보다 높게 나타나, 입상슬러지의 발생기작으로서 polypeptide계 체외폴리머의 존재가능성을 보여주고 있다. 전자현미경을 이용한 형태화적 특성조사결과, 일반적인 반응조의 경우와는 달리 수소분압이 높게 유지된 수정개발된 UASB 반응조의 경우 입상슬러지의 표면에서는 Methanobrevibactor arboriphilus와 같은 크기와 형태를 한 수소이용메탄균의 성장이 다발을 이루며 관찰되었는데, 이러한 현상은 입상슬러지의 형성 메커니즘을 뒷받침해주고 있다. 우수한 입상슬러지의 형성을 위해서는 상대적인 수소분압의 크기에 따른 효과적인 상분리가 이루어져, acetogens과 수소이용메탄균들간의 공생관계가 잘 유지되도록 해주어야 할 것으로 사료된다. 수정개발된 UASB 반응조가 일반적인 UASB 반응조에 비하여 수소이용메탄균의 성장에 더욱 효과적인 환경을 제공하는 것으로 판단되며, 입상슬러지의 경영성과 반응조 전체의 유기물질 제거효율 뿐만 아니라, 운전의 안정성 측면에서도 더 우수한 것으로 사료된다.

• International Journal of Industrial Entomology and Biomaterials
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• 제17권1호
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• pp.125-129
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• 2008

The purpose of this study was to investigate the characteristics of Mamestra brassicae nucleopolyhedrovirus (MabrNPV)-K1 isolated in Korea. Polyhedra of MabrNPV-K1 showed irregular appearance in shape with the average diameter $1.8{\mu}m$. MabrNPV-K1 contained a number of nucleocapsids within a viral envelope embedded in polyhedron. The polyhedrin of MabrNPV-K1 was composed of single polypeptide with a M.W. of approximate 31 kDa which is identical to the commercialized MabrNPV, Mamestrin, as a biological control agent. The nucleotide and amino acid sequences within the coding region of MabrNPV-K1 polyhedrin shared 99.0% similarity with the polyhedrin gene from previous reported MabrNPVs. The median lethal concentrations ($LC_{50}$) of MabrNPV-K1 and Mamestrin to M. brassicae larvae were $3.9{\times}10^3$ PIBs/larva and $6.0{\times}10^4$ PIBs/larva, respectively. Mortality of the MabrNPV-K1 against to the third instars larvae was 15 times higher than that of the Mamestrin. The median lethal times ($LT_{50})$ of MabrNPV-K1 by the concentration of polyhedra were lower ($4.4{\sim}6.1$ days) than those of Mamestrin ($4.1{\sim}8.6$ days). These results suggest that a local strain MabrNPV-K1 has high pathogenicity to M. brassicae and may be useful for the development of biological control agent to control this.

• 한국미생물·생명공학회지
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• 제31권3호
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• pp.216-223
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• 2003

새롭게 분리된 Bacillus cereus H-1으로부터 크기가 45-kDa인 chitosanase를 정제하여 특성을 파악하였고 1.3-kb의 chitosanase 유전자(choA)를 대장균에 클로닝하여 발현시켰다. H-1의 chitosanase 단백질(ChoA)은 ammonium sulfate 침전과 CM-sephadex칼럼 크로마토그래피에 의해 정제하였다. 최적 pH는 약 7이었고 pH 안정성은 $50^{\circ}C$에서 4-11로 나타났다. 최적 온도는 약 5$0^{\circ}C$였으며 효소 활성은 $45^{\circ}C$ 아래에서 비교적 안정하였다. H-1 chitosanase는 soluble 또는 glycol chitosan뿐만아니라 carboxymethyl cellulose(CMC)에 대한 활성도 나타내었다. 정제된 ChoA의 MALDI-TOF MS분석에 기초하여 이미 알려진 다른 Bacillus chitosanases와의 데이터베이스 검색을 통해 전체 아미노산 서열을 밝혀내었다. Chitosanase gene에 해당하는 1.6 kb의 PCR 산물을 얻었으며 그의 DNA 서열을 결정하였다. choA의 추정 아미노산은 Bacillus sp. No 7-M과 Bacillus sp. KCTC0377BP의 아미노산과 98%의 유사성을 나타내었다. 재조합 ChoA단백질은 E. coli DH5$\alpha$에서 원 균주와 동일한 크기로 발현되었다. N말단의 추정아미노산서열을 다른 chitosanas리 서열과 비교해 볼때 ChoA는 chitosanase-cellulase 활성을 갖는 family 8에 속하는 미생물 endo-chitosanaseT. 추정되었다.

• Reproductive and Developmental Biology
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• 제35권3호
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• pp.273-280
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• 2011

It was conducted the experiment, divided into three groups as normal, poor and polycystic ovary syndrome, to detect the change of protein patterns in follicular fluid on ovarian response following controlled ovarian hyperstimulation for human IVF outcome. In the normal group, it was confirmed reproducible 57 spots in the detected total 81 spots. Then 1 spot was not found in the other groups. In the poor responder group, it was found reproducible 53 spots in the detected total 98 spots. 6 spots were down-regulation and 7 spots were up-regulation comparable with normal group. There were not 5 spots in poor responder group comparable with other groups. In the polycystic ovary syndrome group, it was expressed reproducible 53 spots in the detected total 80 spots and 3 spots were just expressed in this group. However, 4 spots were not found in polycystic ovary syndrome. 9 spots were up-regulation comparable with normal group. Significant up and down-regulation spots among the each groups were identified. The results were a cytosolic carboxypeptidase, a signal-induced proliferation-associated protein 1, a ceruloplasmin, a keratin(type II cytoskeletal 1), a polypeptide N-acetylgalactosantinyltransferase 2, a serine/threonine-protein phosphatase 4 regulatory subunit 4. It was identified that 8 spots, 6 kinds of protein are corresponded with NCBInr database research, but 10 spots were failed in the identification. In conclusion, it has been confirmed change and expression of protein on the ovarian response following COH of human.

• Journal of Microbiology and Biotechnology
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• 제25권11호
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• pp.1880-1893
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• 2015

In this study, Pseudomonas R0-14, which was isolated from Arctic soil samples, showed a clear halo when grown on M9 medium agarose plates containing olive oil-rhodamine B as substrate, suggesting that it expressed putative lipase(s). A putative lipase gene, lipR, was cloned from R0-14 by genome walking and Touchdown PCR. lipR encodes a 562-amino-acid polypeptide showing a typical α/β hydrolase structure with a catalytic triad consisting of Ser₁₅₃-Asp₂₀₂-His₂₆₀ and one α-helical lid (residues 103-113). A phylogenetic analysis revealed that LipR belongs to the lipase subfamily I.3. LipR was successfully expressed in Escherichia coli, purified, and biochemically characterized. Recombinant LipR exhibited its maximum activity towards p-nitrophenyl butyrate at pH 8.5 and 60℃ with a K_m of 0.37 mM and a k_cat of 6.42 s^-1. It retained over 90% of its original activity after incubation at 50℃ for 12 h. In addition, LipR was activated by Ca²⁺, Mg²⁺, Ba²⁺, and Sr²⁺, while strongly inhibited by Cu²⁺, Zn²⁺, Mn²⁺, and ethylenediaminetetraacetic acid. Moreover, it showed a certain tolerance to organic solvents, including acetonitrile, isopropanol, acetone, methanol, and tert-butanol. When algal oil was hydrolyzed by LipR for 24 h, there was an enrichment of n-3 long-chain polyunsaturated fatty acids, including eicosapentaenoic acid (1.22%, 1.65-fold), docosapentaenoic acid (21.24%, 2.04-fold), and docosahexaenoic acid (36.98%, 1.33-fold), and even a certain amount of diacylglycerols was also produced. As a result, LipR has great prospect in industrial applications, especially in food and/or cosmetics applications.