• Title/Summary/Keyword: Genetic resource

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Relationship among porcine lncRNA TCONS_00010987, miR-323, and leptin receptor based on dual luciferase reporter gene assays and expression patterns

  • Ding, Yueyun;Qian, Li;Wang, Li;Wu, Chaodong;Li, DengTao;Zhang, Xiaodong;Yin, Zongjun;Wang, Yuanlang;Zhang, Wei;Wu, Xudong;Ding, Jian;Yang, Min;Zhang, Liang;Shang, Jinnan;Wang, Chonglong;Gao, Yafei
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.2
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    • pp.219-229
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    • 2020
  • Objective: Considering the physiological and clinical importance of leptin receptor (LEPR) in regulating obesity and the fact that porcine LEPR expression is not known to be controlled by lncRNAs and miRNAs, we aim to characterize this gene as a potential target of SSC-miR-323 and the lncRNA TCONS_00010987. Methods: Bioinformatics analyses revealed that lncRNA TCONS_00010987 and LEPR have SSC-miR-323-binding sites and that LEPR might be a target of lncRNA TCONS_00010987 based on cis prediction. Wild-type and mutant TCONS_00010987-target sequence fragments and wild-type and mutant LEPR 3'-UTR fragments were generated and cloned into pmiRRB-REPORTTM-Control vectors to construct respective recombinant plasmids. HEK293T cells were co-transfected with the SSC-miR-323 mimics or a negative control with constructs harboring the corresponding binding sites and relative luciferase activities were determined. Tissue expression patterns of lncRNA TCONS_00010987, SSC-miR-323, and LEPR in Anqing six-end-white (AQ, the obese breed) and Large White (LW, the lean breed) pigs were detected by real-time quantitative polymerase chain reaction; backfat expression of LEPR protein was detected by western blotting. Results: Target gene fragments were successfully cloned, and the four recombinant vectors were constructed. Compared to the negative control, SSC-miR-323 mimics significantly inhibited luciferase activity from the wild-type TCONS_00010987-target sequence and wild-type LEPR-3'-UTR (p<0.01 for both) but not from the mutant TCONS_00010987-target sequence and mutant LEPR-3'-UTR (p>0.05 for both). Backfat expression levels of TCONS_00010987 and LEPR in AQ pigs were significantly higher than those in LW pigs (p<0.01), whereas levels of SSC-miR-323 in AQ pigs were significantly lower than those in LW pigs (p<0.05). LEPR protein levels in the backfat tissues of AQ pigs were markedly higher than those in LW pigs (p<0.01). Conclusion: LEPR is a potential target of SSC-miR-323, and TCONS_00010987 might act as a sponge for SSC-miR-323 to regulate LEPR expression.

An Application of a Hybrid Genetic Algorithm on Missile Interceptor Allocation Problem (요격미사일 배치문제에 대한 하이브리드 유전알고리듬 적용방법 연구)

  • Han, Hyun-Jin
    • Journal of the military operations research society of Korea
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    • v.35 no.3
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    • pp.47-59
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    • 2009
  • A hybrid Genetic Algorithm is applied to military resource allocation problem. Since military uses many resources in order to maximize its ability, optimization technique has been widely used for analysing resource allocation problem. However, most of the military resource allocation problems are too complicate to solve through the traditional operations research solution tools. Recent innovation in computer technology from the academy makes it possible to apply heuristic approach such as Genetic Algorithm(GA), Simulated Annealing(SA) and Tabu Search(TS) to combinatorial problems which were not addressed by previous operations research tools. In this study, a hybrid Genetic Algorithm which reinforces GA by applying local search algorithm is introduced in order to address military optimization problem. The computational result of hybrid Genetic Algorithm on Missile Interceptor Allocation problem demonstrates its efficiency by comparing its result with that of a simple Genetic Algorithm.

Isolation and Characterization of Malate Dehydrogenase Gene from Panax ginseng C.A. Meyer (고려인삼에서 Malate Dehydrogenase 유전자의 분리 및 분석)

  • Kim, Yu-Jin;Shim, Ju-Sun;Lee, Jung-Hye;Jung, Dae-Young;In, Jun-Gyo;Lee, Bum-Soo;Min, Byung-Hoon;Yang, Deok-Chun
    • Korean Journal of Medicinal Crop Science
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    • v.16 no.4
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    • pp.261-267
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    • 2008
  • Malate dehydrogenase is a ubiquitous enzyme in plants, involving in a range of metabolic processes depending on its subcellular location. A malate dehydrogenase (PgMDH) cDNA was isolated and characterized from the root of Panax ginseng C. A. Meyer. The deduced amino acid sequence of PgMDH showed high similarity with the NAD-dependent mitochondrial malate dehydrogenase from Glycinemax (P17783), Eucalyptus gunnii (P46487), and Lycopersicon esculentum (AAU29198). And the segment of a malate dehydrogenase gene was amplified through RT-PCR. The expression of PgMDH was increased after treatments of chilling, salt, UV, cadmium or copper treatment.

Relationship between porcine miR-20a and its putative target low-density lipoprotein receptor based on dual luciferase reporter gene assays

  • Ding, Yueyun;Zhu, Shujiao;Wu, Chaodong;Qian, Li;Li, DengTao;Wang, Li;Wan, Yuanlang;Zhang, Wei;Yang, Min;Ding, Jian;Wu, Xudong;Zhang, Xiaodong;Gao, Yafei;Yin, Zongjun
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.7
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    • pp.922-929
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    • 2019
  • Objective: Mutations in low-density lipoprotein receptor (LDLR), which encodes a critical protein for cholesterol homeostasis and lipid metabolism in mammals, are involved in cardiometabolic diseases, such as familial hypercholesterolemia in pigs. Whereas microRNAs (miRNAs) can control LDLR regulation, their involvement in circulating cholesterol and lipid levels with respect to cardiometabolic diseases in pigs is unclear. We aimed to identify and analyze LDLR as a potential target gene of SSC-miR-20a. Methods: Bioinformatic analysis predicted that porcine LDLR is a target of SSC-miR-20a. Wild-type and mutant LDLR 3'-untranslated region (UTR) fragments were generated by polymerase chain reaction (PCR) and cloned into the pGL3-Control vector to construct pGL3 Control LDLR wild-3'-UTR and pGL3 Control LDLR mutant-3'-UTR recombinant plasmids, respectively. An miR-20a expression plasmid was constructed by inserting the porcine premiR-20a-coding sequence between the HindIII and BamHI sites in pMR-mCherry, and constructs were confirmed by sequencing. HEK293T cells were co-transfected with the miR-20a expression or pMR-mCherry control plasmids and constructs harboring the corresponding 3'-UTR, and relative luciferase activity was determined. The relative expression levels of miR-20a and LDLR mRNA and their correlation in terms of expression levels in porcine liver tissue were analyzed using reverse-transcription quantitative PCR. Results: Gel electrophoresis and sequencing showed that target gene fragments were successfully cloned, and the three recombinant vectors were successfully constructed. Compared to pMR-mCherry, the miR-20a expression vector significantly inhibited wild-type LDLR3'-UTR-driven (p<0.01), but not mutant LDLR-3'-UTR-driven (p>0.05), luciferase reporter activity. Further, miR-20a and LDLR were expressed at relatively high levels in porcine liver tissues. Pearson correlation analysis revealed that porcine liver miR-20a and LDLR levels were significantly negatively correlated (r = -0.656, p<0.05). Conclusion: LDLR is a potential target of miR-20a, which might directly bind the LDLR 3'-UTR to post-transcriptionally inhibit expression. These results have implications in understanding the pathogenesis and progression of porcine cardiovascular diseases.

Resource Leveling using Genetic Algorithm (유전알고리즘을 활용한 자원평준화 방법론)

  • Gwak, Han-Seong;Bea, Sang-Hee;Lee, Dong-Eun
    • Journal of the Architectural Institute of Korea Structure & Construction
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    • v.34 no.2
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    • pp.67-74
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    • 2018
  • Resource leveling minimizes resource fluctuations by deferring the earliest start times (ESTs) of non-critical activities within their corresponding total float. The intentional float-consumption for resource leveling purpose reduces the schedule delay contingency. This paper presents a method called Genetic Algorithm based Resource Leveling (GARL) that minimizes resource fluctuations and float-consumption impact over project duration. It identifies activities that are less sensitive to float-consumption and performs resource leveling using those activities. The study is of value to project scheduler because GARL identifies the set of activities to be deferred and the number of shift day(s) of each and every activities in the set within its total float expeditiously. It contributes to establish a baseline schedule which implements an optimal resource leveling plan. A case study is presented to verify the validity and usability of the method. It was confirmed that GARL satisfies the project duration constraint by considering resource fluctuations and float-consumption over project duration.

Conversion of Acidic Polysaccharide and Phenolic Compound of Changed Ginseng by 9 Repetitive Steaming and Drying Process, and Its Effects of Antioxidation (인삼의 구증구포에 의한 산성다당체, 페놀성화합물의 변환 및 항산화능)

  • Kim, Do-Wan;Lee, Yun-Jin;Min, Jin-Woo;Kim, Yu-Jin;Rho, Young-Deok;Yang, Deok-Chun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.1
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    • pp.121-126
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    • 2009
  • Korean ginseng (Panax ginseng C. A. Meyer) has been used as an important medicinal plant in the Orient for a long time. It has been claimed that ginseng has many beneficial bioactive effects on human health, such as antitumor, antistress, antiaging and enhancing immune functions. Red ginseng possibly have new ingredients converted during steaming and dry process from fresh ginseng. In this study, pharmacological efficacy and ingredient conversion of ginseng by 9 repetitive steaming and drying process were investigated measuring conversion efficiency of acidic-polysaccharide, phenolic compounds and inhibition of peroxide lipides. It was found that acidic-polysaccarides were increased by heat treatment. In addition, maltol of phenolic compounds, strong antioxidant, produced during the process of red ginseng by Maillard reaction. Acidic-polysaccarides and maltol were increased after the 1st and 3rd steaming and drying treatments, but they were decreased gradually after 5th, 7th, and 9th treatments. Antioxidant activity was increased as increasing treatment times of steaming and drying without significance. Effect of red ginseng extract on inhibition of peroxide was increased gradually until after the 7th treatment, but remarkably decreased after the 9th treatment.