• Title/Summary/Keyword: M/BP/1

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Chinese Hamster Ovary세포에서 Benzo(a)pyrene과 3-Methylcholanthrene에 의한 DNA 단사절단과 복제억제에 미치는 Saponin의 영향

  • 이정섭;이형호;박기현;박상대
    • Environmental Mutagens and Carcinogens
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    • v.10 no.2
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    • pp.127-134
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    • 1990
  • 본 연구는 Chinese hamster ovary (CHO-KI) 배양세포에서 S-15분획에 의해 활성화된 bezo(a) pyrene (BP)과 3-methylcholanthrene(MC)에 의해 유발된 DNA단사절단과 DNA복제억제 및 그 회복과정에 미치는 한국산 인삼추출물 saponin의 영향을 조사하여 다음과 같은 결과를 얻었다. S-15분획으로 활성화된 10-5 M의 BP 또는 MC를 1.0-10ng/ml의 saponin과 함께 처리할 경우 BP와 MC단독처리군에 비해 DNA 단사절단률이 감소하였다. DNA 합성률은 활성화된 10-5M의 BP와 10-6 M의 MC에 의해 각각 50% 및 75% 억제되었으나, 0.1-10ng/ml의 saponin을 동시에 처리할 경우 DNA 합성억제률이 약 10% 이상 둔화되었다.

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KiSS1, KiSS2, GPR54 mRNA Expression of the Blacktip Grouper Epinephelus fasciatus (홍바리 Epinephelus fasciatus의 KiSS1, KiSS2, GPR54 mRNA 발현양상)

  • Kang, Hyeong-Cheol;Lee, Chi-Hoon;Song, Young-Bo;Baek, Hea-Ja;Kim, Hyung-Bae;Lee, Young-Don
    • Development and Reproduction
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    • v.16 no.2
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    • pp.121-128
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    • 2012
  • Kisspeptin has been reported to facilitate sexual maturation and ovulation by directly stimulating GnRH neurons via its receptor, GPR54. The KiSS-GPR54 system is playing an important role in the reproduction of several mammalian species. However, little is known about their function in fish. The aim of this study is to understand the physiological function and evolutionary conservation of KiSS-GPR54 system in teleost fish blacktip grouper, Epinephelus fasciatus. In the present study, we have partial cloned KiSS1, KiSS2 GPR54 mRNAs from a brain samples. Tissue distribution analysis using RT-PCR revealed that the KiSS1, KiSS2, GPR54 transcripts were expressed in different tissue. The KiSS-GPR54 system in gonadal of immature and mature stage were analyzed using qRT-PCR. The partial sequence of KiSS1, KiSS2, GPR54 were 232 bp, 304 bp, 613 bp long, respectively. KiSS1, KiSS2, GPR54 mRNAs are shown common expression in the brain. The amount of KiSS1, KiSS2 mRNAs expression were significantly higher in mature stage than immature stage. However GPR54 mRNA expression was higher in immature stage. These results are in good agreement with the hypothesis that KiSS-GPR54 system plays an important role in the regulation of reproductive function in the blacktip grouper.

Overexpression and Characterization of Bovine Pancreatic Deoxyribonuclease I in Saccharomyces cerevisiae and Pichia pastoris (Saccharomyces cerevisiae와 Pichia pastoris에서 Bovine Pancreatic Deoxyribonuclease I의 과발현과 특성)

  • Cho, Eun-Soo;Kim, Jeong-Hwan;Yoon, Ki-Hong;Kim, Yeon-Hee;Nam, Soo-Wan
    • Microbiology and Biotechnology Letters
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    • v.40 no.4
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    • pp.348-355
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    • 2012
  • In the present study, we investigated the overexpression and characterization of bovine pancreatic (bp)- DNase I in Saccharomyces cerevisiae and Pichia pastoris. The bp-DNase I gene was fused in frame with the GAL10 promoter, $MF{\alpha}$, and GAL7 terminator sequences, resulting in the plasmid, pGAL-$MF{\alpha}$-DNaseI (6.4 kb). Also, the bp-DNase I gene was fused in frame with the AOX1 promoter, $MF{\alpha}$, and AOX1 terminator sequences, resulting in the plasmid, pPEXI (8.8 kb). The recombinant plasmids, pGAL-$MF{\alpha}$-DNaseI and pPEXI were introduced into S. cerevisiae and P. pastoris host cells, respectively. When the transformed yeast cells were cultured at $30^{\circ}C$ for 48 h in galactose or methanol medium, bp-DNase I was overexpressed and the most of activity was found in the extracellular fraction. P. pastoris transformant activity showed 45.5 unit/mL in the culture medium at 48 h cultivation, whereas S. cerevisiae transformant revealed 37.7 unit/mL in the extracellular fraction at 48 h cultivation. The enzymatic characteristics, such as DNA cleavage and half life were investigated. Treatment of the recombinant DNase I from P. pastoris induced degradation of the calf thymus DNA within 1 minute, and this DNA degradation rate was higher than that of commercial bp-DNase I (SIGMA) and the recombinant DNase I from S. cerevisiae.

The Holocene Depositonal Environment and Sea-Level Change at Ilsan Area (일산충적평야의 홀로세 퇴적환경변화와 해면변동)

  • 황상일
    • Journal of the Korean Geographical Society
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    • v.33 no.2
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    • pp.143-163
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    • 1998
  • In order to clarify the depositional environment and sea-level change at Ilsan area including Gawaji and Saemal valley plains, which is located at the right side in downstream of the Han River, boring data, radiocabon dating and diatom analysis were comprehensively investigated. As a result, the palaeogeographies fo this area altered by the transgressions and regressions after 7,000 y. BP could be restored. The high tide sea-level(mean high water level of spring tide) was arrived ca. 7,000y. BP at the valley plain and risen to ca. 5.5m at ca. 5000y. BP. Since then, the sea-level was kept up the same level to ca. 3,200 BP. The descended sea-level to ca. 2,300 BP was risen up to ca. 5.8m in ca. 1,800 y. BP. It is presumed that such a sea-level change as well as the different sediments in quantity supplied from the river basin of the valley plain could be effected to change diversely the depositional environment of the study area and eventually to the prehistoric human life.

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Characterization of an Easter Lily Calmodulin cDNA Clone (백합실물에서 하나으 Calmodulin cDNA 클론 연구)

  • Kim, Seong-Ryong;An, Gyu-Heung
    • Journal of Plant Biology
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    • v.39 no.1
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    • pp.9-13
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    • 1996
  • A clone, LCM1, which encodes calmodulin (CaM) was isolated and characterized from monocot lily (Lilium longiflorum Thunb.) plants. The clone is 681 bps and contains the 447 bp coding region, 8 bp leader sequence, 210 bp 3'-untraslated region, and a poly(A) tail. The coding region of 149 amino acids encodes a protein of predicted Mr 17 kD. Comparison of the LCM1 amino acid sequence with other CaMs revealed that the protein is highly conserved among various living organisms. The expression level of calmodulin gene in lily was studied by RNA blot analysis. The LCM1 mRNA was present in all tissues tested. However, a higher level of calmodulin was observed in anther and floral bud. The level of calmodulin mRNA in anther was about 10 times higher than that in anther was about 10 times higher than that in vegetative tissues. The anther preferential expression of CaM in lily is currently investigated in dicot plants.

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Substrate Variety of a Non-metal Dependent Tagatose-6-phosphate Isomerase from Staphylococcus aureus (Staphylococcus aureus 유래 비금속성 이성화효소인 Tagatose-6-phosphate Isomerase의 기질다양성)

  • Oh Deok-Kun;Ji Eun-Soo;Kwon Young-Deok;Kim Hye-Jung;Kim Pil
    • Microbiology and Biotechnology Letters
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    • v.33 no.2
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    • pp.106-111
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    • 2005
  • To investigate the substrate variety of a putative non-metal dependent isomerase, the tagatose-6-phosphate isomerase (E.C. 5.3.1.26) structural genes (lacB; 510bp and lacA; 430bp) of Staphylococcus aureus were subcloned and co-expressed. Based on the substrate configuration, various aldoses were surveyed for substrate of ketose isomerization. Among the 10 aldoses tested, D-ribose and D-allose were isomerized by the enzyme. The subunit A and B showed more than $95\%$ activity for D-ribose and $75\%$ for D-allose in the presence of 1mM EDTA compared with non-EDTA conditions, which implying tagatose-6-phosphate isomerase is a non-metal dependent isomerase. Each of subunit A or subunit B alone showed no activity for any of the substrates tested. The affinity constant ($K_m$) of tagatose-6-phosphate isomerase against D-ribose and D-allose were 26 mM and 142 mM, respectively.

Molecular Analysis of Complete SSU to LSU rDNA Sequence in the Harmful Dinoflagellate Alexandrium tamarense (Korean Isolate, HY970328M)

  • Ki, Jang-Seu;Han, Myung-Soo
    • Ocean Science Journal
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    • v.40 no.3
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    • pp.155-166
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    • 2005
  • New PCR primers (N=18) were designed for the isolation of complete SSU to LSU rDNA sequences from the dinoflagellate Alexandrium tamarense. Standard PCR, employing each primer set selected for amplifications of less than 1.5 kb, successfully amplified the expected rDNA regions of A. tamarense (Korean isolate, HY970328M). Complete SSU, LSU rDNAs and ITS sequences, including 5.8S rDNA, were recorded at 1,800 bp, 520 bp and 3,393 bp, respectively. The LSU rDNA sequence was the first report in Alexandrium genus. No intron was found in the LSU rRNA coding region. Twelve D-domains within the LSU rDNA were put together into 1,879 bp (44.4% G+C), and cores into 1514 bp (42.8% G+C). The core sequence was significantly different (0.0867 of genetic distance, 91% sequence similarity) in comparison with Prorocentrum micans (GenBank access. no. X16108). The D2 region was the longest in length (300 bp) and highly variable among the 12 D-domains. In a phylogenetic analysis using complete LSU rDNA sequences of a variety of phytoplankton, A. tamarense was clearly separated with high resolution against other species. The result suggests that the sequence may resolve the taxonomic ambiguities of Alexandrium genus, particularly of the tamarensis complex.

Purification and Characterization of a New Fibrinolytic Enzyme of Bacillus licheniformis KJ-31, Isolated from Korean Traditional Jeot-gal

  • Hwang, Kyung-Ju;Choi, Kyoung-Hwa;Kim, Myo-Jeong;Park, Cheon-Seok;Cha, Jae-Ho
    • Journal of Microbiology and Biotechnology
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    • v.17 no.9
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    • pp.1469-1476
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    • 2007
  • Jeot-gal is a traditional Korean fermented seafood and has long been used for seasoning. We isolated 188 strains from shrimp, anchovy, and yellow corvina Jeot-gal, and screened sixteen strains that showed strong fibrinolytic activities on a fibrin plate. Among those strains, the strain that had the largest halo zone was chosen and identified as Bacillus licheniformis by using 16S rDNA sequencing and an API CHB kit. The fibrinolytic activity of Bacillus licheniformis was characterized and designated as bpKJ-31. The active component of bpKJ-31 was identified as a 37 kDa protein, designated bacillopeptidase F, by internal peptide mapping and N-terminal sequencing. The optimum activity of bpKJ-31 was shown at pH 9 and $40^{\circ}C$, with a chromogenic substrate for plasmin. It had high degrading activity for the $B{\beta}$-chain and $A{\alpha}$-chain of fibrin(ogen), and also acted on thrombin, but not skim milk and casein. The amidolytic activity of bpKJ-31 was inhibited by 1 mM phenylmethanesulfonyl fluoride, but 1 mM EDTA did not affect the enzyme activity, indicating that bpKJ-31 is an alkaline serine protease, like a plasmin. The bpKJ-31 showed approximately 14.3% higher fibrinolytic activity than the plasmin. These features of bpKJ-31 make it attractive as a health-promoting biomaterial.

Amyloid Precursor Protein Binding Protein-1 Is Up-regulated in Brains of Tg2576 Mice

  • Yang, Hyun-Jung;Joo, Yu-Young;Hong, Bo-Hyun;Ha, Sung-Ji;Woo, Ran-Sook;Lee, Sang-Hyung;Suh, Yoo-Hun;Kim, Hye-Sun
    • The Korean Journal of Physiology and Pharmacology
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    • v.14 no.4
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    • pp.229-233
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    • 2010
  • Amyloid precursor protein binding protein-1 (APP-BP1) binds to the carboxyl terminus of amyloid precursor protein and serves as a bipartite activation enzyme for the ubiquitin-like protein, NEDD8. Previously, it has been reported that APP-BP1 rescues the cell cycle S-M checkpoint defect in Ts41 hamster cells, that this rescue is dependent on the interaction of APP-BP1 with hUba3. The exogenous expression of APP-BP1 in neurons has been reported to cause DNA synthesis and apoptosis via a signaling pathway that is dependent on APP-BP1 binding to APP. These results suggest that APP-BP1 overexpression contributes to neurodegeneration. In the present study, we explored whether APP-BP1 expression was altered in the brains of Tg2576 mice, which is an animal model of Alzheimer's disease. APP-BP1 was found to be up-regulated in the hippocampus and cortex of 12 month-old Tg2576 mice compared to age-matched wild-type mice. In addition, APP-BP1 knockdown by siRNA treatment reduced cullin-1 neddylation in fetal neural stem cells, suggesting that APP-BP1 plays a role in cell cycle progression in the cells. Collectively, these results suggest that increased expression of APP-BP1, which has a role in cell cycle progression in neuronal cells, contributes to the pathogenesis of Alzheimer's disease.

Performance improvement of membrane distillation using carbon nanotubes

  • Kim, Seung-Hyun;Lee, Tae-Min
    • Membrane and Water Treatment
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    • v.7 no.4
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    • pp.367-375
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    • 2016
  • Although the bucky paper (BP) made from carbon nanotubes (CNTs) possesses beneficial characteristics of hydrophobic nature and high porosity for membrane distillation (MD) application, weak mechanical strength of BP has often prevented the stable operation. This study aims to fabricate the BP with high mechanical strength to improve its MD performance. The strategy was to increase the purity level of CNTs with an assumption that purer CNTs would increase the Van der Waals attraction, leading to the improvement of mechanical strength of BP. According to this study results, the purification of CNT does not necessarily enhance the mechanical strength of BP. The BP made from purer CNTs demonstrated a high flux ($142kg/m^2{\cdot}h$) even at low ${\Delta}T$ ($50^{\circ}C$ and $20^{\circ}C$) during the experiments of direct contact membrane distillation (DCMD). However, the operation was not stable because a crack quickly formed. Then, a support layer of AAO (anodic aluminum oxide) filter paper was introduced to reinforce the mechanical strength of BP. The support reinforcement was able to increase the mechanical strength, but wetting occurred. Therefore, the mixed matrix membrane (PSf-CNT) using CNTs as filler to polysulphone was fabricated. The DCMD operation with the PSf-CNT membrane was stable, although the flux was low ($6.1kg/m^2{\cdot}h$). This result suggests that the mixed matrix membrane could be more beneficial for the stable DCMD operation than the BP.