• Title/Summary/Keyword: translational efficiency

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Practical Algorisms for PCR-RFLP-Based Genotyping of Echinococcus granulosus Sensu Lato

  • Kim, Hye-Jin;Yong, Tae-Soon;Shin, Myeong Heon;Lee, Kyu-Jae;Park, Gab-Man;Suvonkulov, Uktamjon;Kovalenko, Dmitriy;Yu, Hak Sun
    • Parasites, Hosts and Diseases
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    • v.55 no.6
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    • pp.679-684
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    • 2017
  • Echinococcus granulosus sensu lato (s.l.) is a causative agent of cystic echinococcosis or cystic hydatid disease in humans and domestic and wild animals. The disease is a serious health problem in countries associated with poverty and poor hygiene practices, particularly in livestock raising. We introduced a practical algorism for genotyping the parasite, which may be useful to many developing countries. To evaluate the efficiency of the algorism, we genotyped 3 unknown strains isolated from human patients. We found that unknowns 1 and 3 were included in G1, G2, and G3 genotypes group and unknown 2 was included in G4 genotype (Echinococcus equinus) according to the algorisms. We confirmed these results by sequencing the 3 unknown isolates cox1 and nad1 PCR products. In conclusion, these new algorisms are very fast genotype identification tools that are suitable for evaluating E. granulosus s.l. isolated from livestock or livestock holders, particularly in developing countries.

Characterization of LexA-mediated Transcriptional Enhancement of Bidirectional Hydrogenase in Synechocystis sp. PCC 6803 upon Exposure to Gamma Rays

  • Kim, Jin-Hong;Lee, Min Hee;Kim, Ji Hong;Moon, Yu Ran;Cho, Eun Ju;Kim, Ji Eun;Lee, Choon-Hwan;Chung, Byung Yeoup
    • Rapid Communication in Photoscience
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    • v.1 no.1
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    • pp.21-24
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    • 2012
  • Influence of gamma rays on the cyanobacterium Synechocystis sp. PCC 6803 cells was investigated in terms of a bidirectional hydrogenase, which is encoded by hoxEFUYH genes and responsible for biohydrogen production. Irradiated cells revealed a substantial change in stoichiometry of photosystems at one day after gamma irradiation at different doses. However, as evaluated by the maximal rate of photosynthetic oxygen evolution, maximal photochemical efficiency of photosystem II, and chlorophyll content, net photosynthesis or photosynthetic capacity was not significantly different between the control and irradiated cells. Instead, transcription of hoxE, hoxH, or lexA, which encodes a subunit of bidirectional hydrogenase or the only transcriptional activator, LexA, for hox genes, was commonly enhanced in the irradiated cells. This transcriptional enhancement was more conspicuously observed immediately after gamma irradiation. In contrast, hydrogenase activities were found to somewhat lower in the irradiated cells. Therefore, we propose that transcription of hox genes should be enhanced by gamma irradiation in a LexA-mediated and possibly photosynthesis-independent manner and that this enhancement might not induce a subsequent increase in hydrogenase activities, probably due to the presence of post-transcriptional and/or post-translational regulatory mechanisms.

Enhanced In Vitro Protein Synthesis Through Optimal Design of PCR Primers

  • Ahn Jin-Ho;Son Jeong-Mi;Hwang Mi-Yeon;Kim Tae-Wan;Park Chang-Kil;Choi Cha-Yong;Kim Dong-Myung
    • Journal of Microbiology and Biotechnology
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    • v.16 no.3
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    • pp.355-359
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    • 2006
  • The functional stability of mRNA is one of the crucial factors affecting the efficiency of in vitro translation. As the rapid degradation of mRNA in the cell extract (S30 extract) causes early termination of the translational reactions, extending the mRNA half-life will improve the productivity of the in vitro protein synthesis. Thus, a simple PCR-based method is introduced to increase the stability of mRNA in an S30 extract. The target genes are PCR-amplified with primers designed to make the ends of the transcribed mRNA molecule anneal to each other. When compared with normal mRNA, the mRNA with the annealing sequences resulted in an approximately 2-fold increase of protein synthesis in an in vitro translation reaction. In addition, sequential transcription and translation reactions in a single tube enabled direct protein expression from the PCR-amplified genes without any separate purification of the mRNA.

Codon usage analysis of rice prolamine genes (쌀 저장 단백질 프롤라민 유전자 암호 분석)

  • Lee, Tae-Ho;Kim, Ju-Kon;Nahm, Baek-Hie
    • Applied Biological Chemistry
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    • v.36 no.6
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    • pp.525-532
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    • 1993
  • To characterize the prolamines in rice cultivars, the complete coding sequences of 17 prolamine genes from the database were analyzed. According to the phylogenic analysis of the sequences, these genes could be classified into 4 groups, Group I to IV. The multiple alignment of the deduced amino acid sequences revealed that the four groups differ from one another in chain length caused by deletion of short internal amino acids or carboxyl terminal fragments. Each group was also found to have different amino acid composition with 1, 4, 10 and 30% of sulfur containing amino acids (methionine and cysteine) in Group I to IV prolamines, respectively. Also the isoelectric points of these groups showed the different values of 9.2, 8.2, 6.7 and 7.4. Finally, from the analysis of codon usage pattern of prolamine genes, the codon usage for arginine, serine, threonine, isoleucine, asparagine, aspartic acid, glutamic acid and cysteine were higly biased. In the analysis of the codon usage pattern, the relation of the fraction of G/C ending codons to effective codon numbers suggests the different translational efficiency in the expression of the prolamine multigenes.

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Characterization of the Folding Structure of 3'-end of Lactate Dehydrogenase A-mRNA Isolated from Hormone Stimulated Rat $C_{6}$ Glioma cell culture (홀몬으로 처리된 쥐의 $C_{6}$ glioma 세포배양으로부터 분리된 낙산탈수소 효소 A-mRNA의 3'-말단의 2차 구조)

  • 배석철;이승기
    • Korean Journal of Microbiology
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    • v.25 no.2
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    • pp.94-102
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    • 1987
  • Rat liver LDH A-cDNA has been isolated from a .lambda.gt11-rat lover cDNA library and partially characterized. The size of the isolated rat liver LDH A-cDNA if shown to be 1.6Kb and restriction enzyme sites for the rat liver LDH A-cDNA are also mapped. 682-nucleotide sequence coding for 3'-end of rat liver LDH A-cDNA has been analyzed and compared to the nucleotide sequence of the same region of rat $C_6$-glioma cell LDH A-cDNA which has been cloned from the hormonally stimulated cell cultures. The result shows that 177 nucleotide sequences coding for the C-terminal 59-amino acids are identical but 505 nucleotide sequences of 3'-nontranslated region of the two LSH A-cDNA exhibit characteristic differences in thier nucleotide sequences. Computer analysis for the folding structures for 3'-end 400 nucleotide sequences of the two LDH A-cDNA shows a possibility implying that the two LDH A-mRNAs isolated from different tissues of rats may have different half life and therefore their translational efficiency may be different. It has been previously demonstrated that isoproterenol stimulated rat $C_6$ -glioma cell cultures produce LDH A-mRNA showing 2 to 3-fold longer half life in comparison to that of noninduced LHD A-mRNA. The result therefore support for the idea that hormonally stimulated rat $C_6$-glioma cells may produce LDH A-mRNA containing different nucleotide sequences at the 3'-end nontranslated region by which the hormonally induced LDH A-mRNA could have more stable secondary mRAN structure in comparison to that of noninduced LDH A-mRNA.

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Behavior of Weld Pool Shape and Weld Surface Deformation as a Function of Spot-GTA Welding Position for 304 Stainless Steel (Spot-GTA 용접자세에 따른 304 스테인리스강 용융지 표면 및 용접부 형상 거동)

  • Kang, Nam-Hyun;Park, Yeong-Do;Cho, Kyung-Mox;Singh, Jogender;Kulkarni, Anil
    • Journal of Welding and Joining
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    • v.26 no.2
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    • pp.62-68
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    • 2008
  • Effects of gravitational orientation on gas tungsten arc welding (GTAW) for 304 stainless steel were studied to determine the critical factors for weld pool formation, such as weld surface deformation and weld pool shape. This study was accomplished through an analytical study of weld pool stability as a function of primary welding parameters (arc current and arc holding time), material properties (surface tension and density), and melting efficiency (cross-sectional area). The stability of weld pool shape and weld surface deformation was confirmed experimentally by changing the welding position. The arc current and translational velocity were the major factors in determining the weld pool stability as a function of the gravitational orientation. A 200A spot GTAW showed a significant variation of the weld pool formation as the arc held longer than 3 seconds, however the weld pool shape and surface morphology for a 165A spot GTAW were 'stable', i.e., constant regardless of the gravitational orientation. The cross-sectional area of the weld (CSA) was one of the critical factors in determining the weld pool stability. The measured CSA ($13.5mm^2$) for the 200A spot GTAW showed a good agreement with the calculated CSA ($14.9mm^2$).

Efficacy of Recombinant Baculovirus Vector Reconstructed in pcDNA3.1 Vector (pcDNA3.1 벡터에서 재구성된 재조합 Baculovirus 벡터의 효능)

  • Sa, Young-Hee;Choi, Chang-Shik;Lee, Ki Hwan;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2018.10a
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    • pp.444-447
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    • 2018
  • Baculovirus expression systems have many known advantages including fast and cost-effective methods to generate large amounts of recombinant proteins in comparison to bacterial expression systems, particularly those requiring complex post-translational modifications. Especially, recombinant baculoviruses can transfer their vectors and express their recombinant proteins in a wide range of mammalian cell types. In this study, baculoviral vectors which were reconstructed from pcDNA3.1 vector, were recombined with cytomegalovirus (CMV) promoter,uroplakin II promoter, polyhedron promoter, vesicular stomatitis virus G (VSVG), enhanced green fluorescent protein (EGFP), and protein transduction domain (PTD). These recombinant vectors were infected with various cells and cell lines. The baculovirus vector thus developed was analyzed by comparing the metastasis and expression of the recombinant genes with conventional vectors. These results suggest that the baculovirus vector has higher efficiency in metastasis and expression than the control vector.

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Effect of Wnt signaling pathway activation on the efficient generation of bovine intestinal organoids

  • Park, Kang Won;Yang, Hyeon;Wi, Hayeon;Ock, Sun A;Lee, Poongyeon;Hwang, In-Sul;Lee, Bo Ram
    • Journal of Animal Reproduction and Biotechnology
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    • v.37 no.2
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    • pp.136-143
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    • 2022
  • Recent progress has been made to establish intestinal organoids for an in vitro model as a potential alternative to an in vivo system in animals. We previously reported a reliable method for the isolation of intestinal crypts from the small intestine and robust three-dimensional (3D) expansion of intestinal organoids (basal-out) in adult bovines. The present study aimed to establish next-generation intestinal organoids for practical applications in disease modeling-based host-pathogen interactions and feed efficiency measurements. In this study, we developed a rapid and convenient method for the efficient generation of intestinal organoids through the modulation of the Wnt signaling pathway and continuous apical-out intestinal organoids. Remarkably, the intestinal epithelium only takes 3-4 days to undergo CHIR (1 µM) treatment as a Wnt activator, which is much shorter than that required for spontaneous differentiation (7 days). Subsequently, we successfully established an apical-out bovine intestinal organoid culture system through suspension culture without Matrigel matrix, indicating an apical-out membrane on the surface. Collectively, these results demonstrate the efficient generation and next-generation of bovine intestinal organoids and will facilitate their potential use for various purposes, such as disease modeling, in the field of animal biotechnology.

The effects of Korean Red Ginseng-derived components on oligodendrocyte lineage cells: Distinct facilitatory roles of the non-saponin and saponin fractions, and Rb1, in proliferation, differentiation and myelination

  • Lee, Ahreum;Kwon, Oh Wook;Jung, Kwi Ryun;Song, Gyun Jee;Yang, Hyun-Jeong
    • Journal of Ginseng Research
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    • v.46 no.1
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    • pp.104-114
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    • 2022
  • Background: Abnormalities of myelin, which increases the efficiency of action potential conduction, are found in neurological disorders. Korean Red Ginseng (KRG) demonstrates therapeutic efficacy against some of these conditions, however effects on oligodendrocyte (OL)s are not well known. Here, we examined the effects of KRG-derived components on development and protection of OL-lineage cells. Methods: Primary OL precursor cell (OPC) cultures were prepared from neonatal mouse cortex. The protective efficacies of the KRG components were examined against inhibitors of mitochondrial respiratory chain activity. For in vivo function of Rb1 on myelination, after 10 days of oral gavage into adult male mice, forebrains were collected. OPC proliferation were assessed by BrdU incorporation, and differentiation and myelination were examined by qPCR, western blot and immunocytochemistry. Results: The non-saponin promoted OPC proliferation, while the saponin promoted differentiation. Both processes were mediated by AKT and extracellular regulated kinase (ERK) signaling. KRG extract, the saponin and non-saponin protected OPCs against oxidative stress, and both KRG extract and the saponin significantly increased the expression of the antioxidant enzyme. Among 11 major ginsenosides tested, Rb1 significantly increased OL membrane size in vitro. Moreover, Rb1 significantly increased myelin formation in adult mouse brain. Conclusion: All KRG components prevented OPC deaths under oxidative stress. While non-saponin promoted proliferation, saponin fraction increased differentiation and OL membrane size. Furthermore, among all the tested ginsenosides, Rb1 showed the biggest increase in the membrane size and significantly enhanced myelination in vivo. These results imply therapeutic potentials of KRG and Rb1 for myelin-related disorders.

Geometrical Design and SLIPS Lubrication for Enhancement of Negative-pressure-driven Internal Flow Rate in Metal Pipes (금속관 내부의 음압유량 향상을 위한 기하학적 디자인 및 SLIPS 윤활)

  • Kim, Dong Geun;Jang, Changhwan;Kim, Seong Jae;Kim, Daegyoum;Kim, Sanha
    • Tribology and Lubricants
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    • v.37 no.6
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    • pp.253-260
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    • 2021
  • Metal pipes are used in a wide range of applications, from plumbing systems of large construction sites to small devices such as medical tools. When a liquid is enforced to flow through a metal pipe, a higher flow rate is beneficial for higher efficiency. Using high pressures can enhance the flow rate yet can be harmful for medical applications. Thus, we consider an optimal geometrical design to increase the flow rate in medical devices. In this study, we focus on cannulas, which are widely used small metal pipes for surgical procedures, such as liposuction. We characterize the internal flow rate driven by a negative pressure and explore its dependence on the key design parameters. We quantitatively analyze the suction characteristics for each design variable by conducting computational fluid dynamics simulations. In addition, we build a suction performance measurement system which enables the translational motion of cannulas with pre-programmed velocity for experimental validation. The inner diameter, section geometry, and hole configuration are the design factors to be evaluated. The effect of the inner diameter dominates over that of section geometry and hole configuration. In addition, the circular tube shape provides the maximum flow rate among the elliptical geometries. Once the flow rate exceeds a critical value, the rate becomes independent of the number and width of the suction holes. Finally, we introduce a slippery liquid-infused nanoporous surface (SLIPS) coating using nanoparticles and hydrophobic lubricants that effectively improves the flow rate and antifouling property of cannulas without altering the geometrical design parameter.