• Title/Summary/Keyword: reverse tolerance

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Changes in Stanniocalcin-2 and Hypoxia-Inducible Factor-1α mRNA Expression in Medaka Oryzias dancena Exposed to Acute Hypoxia (저산소환경에 의한 송사리(Oryzias dancena)의 Stanniocalcin-2와 Hypoxia-Inducible Factor-1α mRNA 발현의 변화)

  • Shin, Ji Hye;Sohn, Young Chang
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.46 no.1
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    • pp.70-76
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    • 2013
  • Some fish live in aquatic environments with low or temporally changing $O_2$ availability. Variation in dissolved oxygen (DO) levels requires behavioral, physiological, and biochemical adaptations to ensure the uptake of sufficient $O_2$. Several species are relatively well adapted to tolerate low $O_2$ partial pressures (hypoxia). The medaka (Oryzias dancena ) is an important model organism for biomedical research that shows remarkable tolerance to hypoxia. We investigated the regulation and role of hypoxia-inducible factor-1 (HIF-$1{\alpha}$) as a general hypoxia-response gene and stanniocalcin-2 (STC2), which is one of the genes regulated by HIF-$1{\alpha}$ in mammals under hypoxia. We subjected adult male medaka to the following three acute hypoxia regimes: 1, 24, and 72 h at DO = $1.8{\pm}0.5$ ppm. The changes in STC2 and HIF-$1{\alpha}$ mRNA were monitored using quantitative real-time reverse-transcription PCR. We found strong upregulation of HIF-$1{\alpha}$ mRNA in the livers of fish exposed to hypoxia. Hypoxia rapidly upregulated STC-2 mRNA expression in muscle, but not in the brain, gills, liver, or intestine. Therefore, unlike in mammals, hypoxia might regulate O. dancena STC-2 expression in an HIF-$1{\alpha}$-independent manner.

Effective Exon-Intron Structure Verification of a 1-Pyrroline-5-Carboxylate-Synthetase Gene from Halophytic Leymus chinensis (Trin.) Based on PCR, DNA Sequencing, and Alignment

  • Sun, Yan-Lin;Hong, Soon-Kwan
    • Korean Journal of Plant Resources
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    • v.23 no.6
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    • pp.526-534
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    • 2010
  • Genomes of clusters of related eukaryotes are now being sequenced at an increasing rate. In this paper, we developed an accurate, low-cost method for annotation of gene prediction and exon-intron structure. The gene prediction was adapted for delta 1-pyrroline-5-carboxylate-synthetase (p5cs) gene from China wild-type of the halophytic Leymus chinensis (Trin.), naturally adapted to highly-alkali soils. Due to complex adaptive mechanisms in halophytes, more attentions are being paid on the regulatory elements of stress adaptation in halophytes. P5CS encodes delta 1-pyrroline-5-carboxylate-synthetase, a key regulatory enzyme involved in the biosynthesis of proline, that has direct correlation with proline accumulation in vivo and positive relationship with stress tolerance. Using analysis of reverse transcription-polymerase chain reaction (RT-PCR) and PCR, and direct sequencing, 1076 base pairs (bp) of cDNA in length and 2396 bp of genomic DNA in length were obtained from direct sequencing results. Through gene prediction and exon-intron structure verification, the full-length of cDNA sequence was divided into eight parts, with seven parts of intron insertion. The average lengths of determinated coding regions and non-coding regions were 154.17 bp and 188.57 bp, respectively. Nearly all splice sites displayed GT as the donor sites at the 5' end of intron region, and 71.43% displayed AG as the acceptor sites at the 3' end of intron region. We conclude that this method is a cost-effective way for obtaining an experimentally verified genome annotation.

Molecular Cloning of Hemoglobin Alpha-chain Gene from Pantholops hodgsonii, a Hypoxic Tolerance Species

  • Yingzhong, Yang;Droma, Yunden;Guoen, Jin;Zhenzhong, Bai;Lan, Ma;Haixia, Yun;Yue, Cao;Kubo, Keishi;Rili, Ge
    • BMB Reports
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    • v.40 no.3
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    • pp.426-431
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    • 2007
  • To investigate the possible mechanisms of high-altitude native animals in adapting to high altitude, we cloned hemoglobin alpha-chain (alpha-chain Hb) gene from Pantholops hodgsonii, an animal species that indigenously lives at elevations of 3700-5500 m on the Qinghai-Tibetan plateau. Using reverse transcription polymerase chain reaction (RT-PCR) technique, the alpha-chain Hb gene was amplified from total RNA in the liver of the Pantholops hodgsonii. TA cloning technique was used and the PCR product was cloned into pGEM-T vector. The DNA sequence of the gene was highly homologous with sheep (99.1%), goat (98.6%), cattle (95.6%) and human (86.5%). The alpha-chain Hb gene encoded a 142-amino acid protein that could be identified with the homology of alpha-chain Hb protein in sheep (98%), goat (96%), cattle (91%) and human (87%). However, 18 alternations were detected when compared with the alpha-chain Hb gene in human, and 2 in sheep. Moreover, the alterations of a117 GluAsp and $\alpha$132 AsnSer in important regions were noted in human and sheep, respectively. Phylogenetic analysis suggested that the structure of alpha-chain Hb was highly similar to that in sheep. This study provided essential information for elucidating the possible roles of hemoglobin in adapting to extremely high altitude in Pantholops hodgsonii.

An Efficient Plant Regeneration and Transformation System of Robinia pseudoacacia var. umbraculifera for Phytoremediation

  • Kwon, Hye-Jin;Woo, Seong-Min;Seul, Eun-Jun;Kim, Teh-Ryung;Shin, Dong-Un;Kim, Hag-Hyun
    • Journal of Plant Biotechnology
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    • v.34 no.4
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    • pp.293-298
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    • 2007
  • Robinia pseudoacacia var. umbraculifera, commonly called umbrella black locust were regenerated after co-cultivation of internode segments with Agrobacterium tumefaciens which included yeast cadmium factor 1 (YCF 1) gene. The tolerance to cadmium and lead for plants can be increased by the YCF1 gene expression. Moreover, the recent studies have shown that YCF1 gene transgenic plants increase the accumulation of cadmium and lead into plant vacuoles. The effect of plant growth regulator such as 2,4-dichlorophenoxyacetic acid (2,4-D), ${\alpha}$-naphthaleneacetic acid (NAA), 6-benzyladenine (BA), and thidiazuron (TDZ) were studied to evaluate the propagation of plants through internode explants. The efficient induction of multiple adventitious shoots and callus were observed on a medium supplemented with 0.1 mg/L TDZ + 0.2 mg/L BA. To induce shoot elongation and rooting, regenerated shoots were transferred into basal MS medium without any plant growth regulator. Successful Agrobacterium tumefaciens mediated transformation was obtained by 20 min vacuum-infiltration with $50{\mu}M$ acetosyringone on the optimal multiple shoot induction medium with 30 mg/L hygromycin and 300 mg/L cefotaxime. To confirm the integration and expression of transgene, Polymerase Chain Reaction (PCR) and Reverse Transcriptase PCR (RT-PCR) were performed with specific primers. The frequency of transformation was approximately 18.94%. This study can be used to genetic engineering of phytoremediator.

Identification of Copper and Cadmium Induced Genes in Alfalfa Leaves through Annealing Control Primer Based Approach

  • Lee, Ki-Won;Rahman, Md. Atikur;Zada, Muhammad;Lee, Dong-Gi;Kim, Ki-Yong;Hwang, Tae Young;Ji, Hee Jung;Lee, Sang-Hoon
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.35 no.3
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    • pp.264-268
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    • 2015
  • The present research investigated copper and cadmium stress-induced differentially expressed genes (DEGs) using annealing control primers (ACP) with the differential display reverse transcription polymerase chain reaction technique in alfalfa (Medicago sativa L. cv. Vernal) leaves. Alfalfa leaves were subjected to $250{\mu}M$ of copper and cadmium treatment for a period of 6 h. A total of 120 ACPs was used. During copper and cadmium treatment, 6 DEGs were found to be up or down regulated. During copper stress treatment, 1 DEG was up-regulated, and 3 novel genes were discovered. Similarly, during cadmium stress treatment, 1 DEG was up-regulated and 5 novel genes were identified. Among all 6 DEGs, DEG-4 was identified as the gene for trans-2,3-enoyl-CoA reductase, DEG-5 was identified as the gene for senescence-associated protein DIN1 and DEG-6 was identified for caffeic acid O-methyltransferase. All the up-regulated genes may play a role in copper and cadmium stress tolerance in alfalfa.

The effect of Korean Red Ginseng extract on rosiglitazone-induced improvement of glucose regulation in diet-induced obese mice

  • Oh, Mi-Jeong;Kim, Hyun-Ju;Park, Eun-Young;Ha, Na-Hee;Song, Mun-Gyu;Choi, Sang-Hyun;Chun, Boe-Gwun;Kim, Dong-Hoon
    • Journal of Ginseng Research
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    • v.41 no.1
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    • pp.52-59
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    • 2017
  • Background: Korean Red Ginseng extract (KRG, Panax ginseng Meyer) and its constituents have been used for treating diabetes. However, in diet-induced obese mice, it is unclear whether KRG can enhance the glucose-lowering action of rosiglitazone (ROSI), a peroxisome proliferator-activated receptor gamma synthetic activator. Methods: Oral glucose tolerance tests (oGTTs) were performed after 4 days of treatment with a vehicle (CON), KRG [500 mg/kg body weight (b.w.)], ROSI (3.75 mg/kg b.w, 7.5 mg/kg b.w, and 15 mg/kg b.w.), or ROSI and KRG (RK) in obese mice on a high-fat diet. Adipose tissue morphology, crown-like structures (CLSs), and inflammation were compared by hematoxylin-eosin staining or quantitative reverse transcription polymerase chain reaction. Results: The area under the glucose curve (AUC) was significantly lower in the RK group (15 mg/kg b.w. and 500 mg/kg b.w. for ROSI and KRG, respectively) than in the CON group. There was no significant difference in the AUC between the CON and the other groups. Furthermore, the AUC was significantly lower in the RK group than in the ROSI group. The expression of the Ccl2 gene and the number of CLSs were significantly reduced in the RK group than in the CON group. Conclusion: Our results show a potential enhancement of ROSI-induced improvement of glucose regulation by the combined treatment with KRG.

Physiological Response to Salinity Stress of Japonica/Indica Lines Tolerant to Salt at Seedling Stage

  • Ko, Jong-Cheol;Lee, Kyu-Seong;Kim, Ki-Young;Choi, Weon-Young;Kim, Bo-Kyung;Shin, Woon-Cheol;Ko, Jae-Kwon;Yum, Song-Joong
    • Korean Journal of Breeding Science
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    • v.43 no.5
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    • pp.391-398
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    • 2011
  • Physiological responses to salinity stress were evaluated in six rice genotypes differing in their tolerance to salinity at the seedling stage. Susceptible genotypes ('Dongjingbyeo', 'Hwayeongbyeo', and 'IR29') showed salt injury symptoms (mean 8.8) and higher visual score under salt stress than that of tolerant ones ('Pokkali', 'IR74009', and 'IR73571'). As salinity affects growth and physiological parameters, the six genotypes thus showed significant reduction because of salt stress. Tolerant Japonica/Indica bred lines ('IR74009', 'IR73571') showed lower reduction, 33.9%, 34.5%, and 50%, respectively, in plant seedling height, dry shoot weight and dry root weight than those of the susceptible Japonica varieties ('Dongjingbyeo', 'Hwayeongbyeo'), and the highest reduction under salt stress was observed in dry root weight, followed by dry shoot weight and seedling height, respectively. Shoot $Na^+$ concentration of IR74099 and IR73571 was lower than that of the susceptible varieties, 'Dongjinbyeo' and 'Hwayeongbyeo'. There were no significant differences among genotypes in root $Na^+$ concentration. Shoot $K^+$ concentration showed a reverse tendency compared to shoot $Na^+$ concentration. IR74009 and IR73571 had considerably lower ratio compared to 'Dongjinbyeo' and 'Hwayeongbyeo' in $Na^+/K^+$ ratio of their shoot and was not different the tolerant check, 'Pokkali'.

Modeling Technology on Free-form Surface of a New Military Personal Head using Quick Surface Method (퀵서피스기법을 이용한 신장병 두상의 자유곡면 모델링 기술)

  • Lee, Yong-Moon;Hwang, Tae-Son;Kim, Hun;Nam, Hee-Tae;Lee, Kee-Hwan;Kang, Myungchang
    • Journal of the Korean Society of Manufacturing Process Engineers
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    • v.17 no.6
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    • pp.170-176
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    • 2018
  • Recently, weapon system requires personal protection products due to the explosion of rapid-fire explosion, which is considered to be multi threat in modernization, complication and war against terrorism. However, the conventional Korean military bullet protection helmets are not suitable for wearing convenience and combatant interoperability in terms of ergonomic. In this paper, we propose a suitable 3D Scanning method for the head, and compare the measured 3D dimension with the existing 2D measurement value to identity the reliability. Reverse engineered soldier head using the quick surface method was realized with a perfect free-form surface and satisfactory tolerance range (${\pm}0.2mm$). Through the comparison of 3D and 2D measured head dimensions, the absolute error value was 0.73 mm on average and relative error was 0.35 %, confirming the high accuracy of the 3D scan modeling. Also, quick surface method using 3D scanner is suggested a fast and accurate skill for ergonomics in obtaining the head modeling needed for military's personal bullet protection helmet design.

NaCl Concentration-Dependent Aminoglycoside Resistance of Halomonas socia CKY01 and Identification of Related Genes

  • Park, Ye-Lim;Choi, Tae-Rim;Kim, Hyun Joong;Song, Hun-Suk;Lee, Hye Soo;Park, Sol Lee;Lee, Sun Mi;Kim, Sang Hyun;Park, Serom;Bhatia, Shashi Kant;Gurav, Ranjit;Sung, Changmin;Seo, Seung-Oh;Yang, Yung-Hun
    • Journal of Microbiology and Biotechnology
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    • v.31 no.2
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    • pp.250-258
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    • 2021
  • Among various species of marine bacteria, those belonging to the genus Halomonas have several promising applications and have been studied well. However, not much information has been available on their antibiotic resistance. In our efforts to learn about the antibiotic resistance of strain Halomonas socia CKY01, which showed production of various hydrolases and growth promotion by osmolytes in previous study, we found that it exhibited resistance to multiple antibiotics including kanamycin, ampicillin, oxacillin, carbenicillin, gentamicin, apramycin, tetracycline, and spectinomycin. However, the H. socia CKY01 resistance pattern to kanamycin, gentamicin, apramycin, tetracycline, and spectinomycin differed in the presence of 10% NaCl and 1% NaCl in the culture medium. To determine the mechanism underlying this NaCl concentration-dependent antibiotic resistance, we compared four aminoglycoside resistance genes under different salt conditions while also performing time-dependent reverse transcription PCR. We found that the aph2 gene encoding aminoglycoside phosphotransferase showed increased expression under the 10% rather than 1% NaCl conditions. When these genes were overexpressed in an Escherichia coli strain, pETDuet-1::aph2 showed a smaller inhibition zone in the presence of kanamycin, gentamicin, and apramycin than the respective control, suggesting aph2 was involved in aminoglycoside resistance. Our results demonstrated a more direct link between NaCl and aminoglycoside resistance exhibited by the H. socia CKY01 strain.

Generation of Transgenic Mice Overexpression Mouse RESISTIN

  • J. R. Chun;S. J. Song;J. T. Do;K. S. Chung;Lee, H. T.
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.99-99
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    • 2002
  • The hormone resistin is associated with typeII diabetes mellitus in rodent model. Resistin impairs glucose tolerance and insulin action. A new class of anti-diabetic drugs were called thiazolidinediones (TZDs) downregulates a resistin which is induced during adipocyte differentiation. But the connection between increased adiposity and resistin remains unknown. The objectives of this study was to clone a mouse resistin cDNA and to generate transgenic mice overexpressing mouse resistin gene. The 555 bp of mouse resistin was amplified from mob cDNAS by polymerase chain reaction (PCR) and cloned into pCR$\^$(R)/ 2.1 TOPO T-vector. Mouse resistin mRNA on the basis of Genbank sequence (acession no. AF323080). Then, the PCR product was cloned into pTargeT$\^$TM/ mammalian expression vector that has pCMV promoter and chimeric intron. Restriction enzyme analysis with BamH I and Not I was carried out to determine an orientation of the insert in the vector. The pCMV-mus/resistin gene was prepared from previous recombinant pTargeT$\^$TM/-mus/resistin by digestion of Bgl II, and has used for microinjection into pronuclei of one cell embryos. The microinjected embryos were transfered to pseudopregnant foster-mother. Mouse resistin expression was detected in transgenic F1 mice by Reverse Transcriptase- Polymerase Chain Reaction (RT-PCR). Resistin gene expression mouse has heavier body weight which was measured higher level of plasma glucose than that of normal mouse. And in diet-induced experiments, the abdominal fat pads were isolated from each 24h starvation and re-feeding after fasting group mice that were assessed by RT-PCR analysis. In fasting group mice, resistin expression was higher than that of re-feeding group mice. This result suggests that the resistin gene overexpressing mice may be became to obesity and be useful as an animal disease model to be diabetes mellitus caused by insulin resistance of resistin.

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