• Title/Summary/Keyword: Resistant mutant

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Breeding of Self-compatible Pear "Wonkyo Na-jasoojung 2" (배 자가결실성 "원교 나-자수정 2호")

  • Shin, Il Sheob;Shin, Yong Uk;Hwang, Hae Sung;Heo, Seong;Kim, Ki Hong;Kang, Sam Seok;Kim, Yoon Kyeong
    • Korean Journal of Breeding Science
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    • v.41 no.2
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    • pp.154-157
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    • 2009
  • Pear has a gametophytic self-incompatibility (SI) system and its SI reaction is controlled by a single multi-allelic S-locus. 'Wonkyo Na-jasoojung 2' was selected from a cross between 'Wonwhang', early season major pear cultivar with high fruit quality and self-incompatible, and 92-18-79 (${S_4}^{sm}{S_4}^{sm}$) obtained from self cross of 'Osa-nijisseiki' (${S_2S_4}^{sm}$) (SM, stylar-part mutant), self-compatible bud mutant that originated from self-incompatible 'Nijisseiki' ($S_2S_4$) made in 2001 at the National Institute of Horticultural and Herbal Science, Rural Development Administration in Korea. '92-18-79' was selected as a self-compatible source through field investigation. It bloomed 1 day earlier than 'Osa-Nijisseiki' and similar to 'Wonwhang' in 2008. It is medium in tree vigor and spreading in tree habit. 'Wonkyo Na-jasoojung 2' is classified as highly susceptible to pear scab (Venturia nashicola) similar to 'Osa-Nijisseiki' and as resistant to black spot (Alternaria kikuchiana) similar to 'Wonwhang'. It had 65.7% fruiting rate by self pollination. The average optimum harvest time of 'Wonkyo Na-jasoojung 2' was 148 days after full bloom and it matured 2 days earlier than 'Osa-Nijisseiki' and 11 days later than 'Wonwhang'. The fruit is roundish oblate in shape and yellowish brown in skin color. Average fruit weight was 445 g and soluble solids content was 13.3 $^{\circ}Brix$. The flesh had abundant juice and negligible grit.

Effects of Coinoculation by Bradyrhizobium japonicum with other Rhizobacteria and of Antibiotic Treatment on Soybean Growth and Nodule Formation (근류균(根瘤菌)과 타(他) 근권미생물(根圈微生物)과의 혼합접종(混合接種) 및 항생제처리(抗生劑處理)가 대두(大豆)의 생장(生長)과 근류형성(根瘤形成)에 미치는 영향(影響))

  • Park, Byeong-Jun;Kim, Yong-Woong;Kim, Kwang-Sik
    • Korean Journal of Soil Science and Fertilizer
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    • v.29 no.2
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    • pp.190-198
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    • 1996
  • The Neomycin resistant gene in transposon 5 (Tn5) was introduced into a mutant of Bradyrhizobium japonicum RJB 6 $str^rnal^r$ by conjugation. This marked strain was used in coinoculation of soybean with Pseudomonas fluorescens(Ps-3) and Bacillus subtilis(BCAC-4) and in antibiotic treatment of soybean for studying rate of nodule formation and growth promoting effect on soybean plant. When the marked strain RJB 6 $str^rnal^rneo^r$ with Ps-3, BCAC-4, and Ps-3 plus BCAC-4 was coinoculated into two soils, the recovery rate of the marked strain was 8.5, 4.3 and 2.9 percent in soybean cultivated soil, and 10.3. 6.0 and 5.0 percent in soybean uncultivated soil. The best growth promoting effect of coinoculation on soybean plant was found with the marked strain plus Ps-3. When the marked strain was inoculated into soybean with antibiotic treatment, the rate of nodule formation in soybean cultivated soil was between 12.5 and 25.4 percent, while that in soybean uncultivated soil was between 23.7 and 43.2 percent. The highest rate of nodule formation with the marked strain was found in streptomycin 1000 ppm treatment plot.

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Isolation, Identification and Mutant Development of Butanol Tolerance Bacterium (부탄올 내성 미생물의 분리, 동정 및 변이주의 개발)

  • Jung, Hyesook;Lee, Jinho
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.26-32
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    • 2013
  • Butanol-resistant bacteria were isolated from butanol solvent. The cell growth of isolated strains declined with increasing concentrations of butanol, and isolated strain BRS02 displayed more resistance to 12.5 g/L of butanol than other isolated strains. In addition, strain BRS251, which was resistant to even higher concentrations of butanol, was developed by the mutation of BRS02 using UV. BRS251 could grow in LB medium containing up to 17.5 g/L of butanol, 32.5 g/L of propanol, or 6 g/L of pentanol, whereas the control strain Escherichia coli was found to be tolerant to 7.5 g/L of butanol, 20 g/L of propanol, or 2 g/L of pentanol. The isolated BRS02, a Gram(+) bacterium seen to have a cocci form under the microscope, grew in 6.5% NaCl. According to biochemical tests, BRS02 can metabolize and produce acid with D-galactose, D-maltose, D-mannitol, D-mannose, methyl-${\beta}$-Dglucopyranoside, D-ribose, sucrose, or D-trehalose, as carbon sources. Also, this strain showed resistance to bacitracin, vibriostatic agent O/129, and optochin, alongside positive activities for arginine dihydrolase, ${\alpha}$-glucosidase, and urease. The BRS02 strain was identified as Staphylococcus sp. by analyses of the 16S rRNA gene, phylogenetic tree, and biochemical tests.

Snail Switches 5-FU-induced Apoptosis to Necrosis through Akt/PKB Activation and p53 Down-regulation (Snail의 Akt/PKB의 활성화와 p53의 downregulation를 통한 5-FU-induced apoptosis의 necrosis로의 전환)

  • Lee, Su-Yeon;Jeon, Hyun-Min;Ju, Min-Kyung;Kim, Cho-Hee;Jeong, Eui-Kyong;Park, Hye-Gyeong;Kang, Ho-Sung
    • Journal of Life Science
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    • v.22 no.8
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    • pp.1018-1023
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    • 2012
  • Snail is a zinc finger transcription factor that induces epithelial-to-mesenchymal transition (EMT), which promotes tumor invasion and metastasis by repressing E-cadherin expression. In addition, Snail restricts the cellular apoptotic response to apoptotic stimuli or survival factor withdrawal; however, its molecular mechanism remains largely unknown. In this study, we have investigated the mechanism underlying Snail-mediated chemoresistance to 5-fluorouracil (5-FU), one of the most widely used anti-cancer drugs. When Snail was overexpressed by doxycycline (DOX) in MCF-7 #5 cells, it inhibited 5-FU-induced apoptotic cell death and switched the cell death mode to necrosis. Snail expression, either by DOX treatment in MCF-7 #5 cells or by the transfection of Snail expression vectors pCR3.1-Snail-Flg, phosphorylation-resistant pCR3.1-S104, and 107A Snail-Flg in MCF-7 cells specifically induced PTEN down-regulation/inactivation and Akt/PKB activation, without affecting ERK1/2 activity. In addition, Snail prominently suppressed 5-FU-induced increases in p53 levels. These findings demonstrate that Snail switches 5-FU-induced apoptosis to necrosis through the activation of Akt/PKB and the down-regulation of p53 levels.

Mutations in the gyrB, parC, and parE Genes of Quinolone-Resistant Isolates and Mutants of Edwardsiella tarda

  • Kim, Myoung-Sug;Jun, Lyu-Jin;Shin, Soon-Bum;Park, Myoung-Ae;Jung, Sung-Hee;Kim, Kwang-Il;Moon, Kyung-Ho;Jeong, Hyun-Do
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1735-1743
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    • 2010
  • The full-length genes gyrB (2,415 bp), parC (2,277 bp), and parE (1,896 bp) in Edwardsiella tarda were cloned by PCR with degenerate primers based on the sequence of the respective quinolone resistance-determining region (QRDR), followed by elongation of 5' and 3' ends using cassette ligation-mediated PCR (CLMP). Analysis of the cloned genes revealed open reading frames (ORFs) encoding proteins of 804 (GyrB), 758 (ParC), and 631 (ParE) amino acids with conserved gyrase/topoisomerase features and motifs important for enzymatic function. The ORFs were preceded by putative promoters, ribosome binding sites, and inverted repeats with the potential to form cruciform structures for binding of DNA-binding proteins. When comparing the deduced amino acid sequences of E. tarda GyrB, ParC, and ParE with those of the corresponding proteins in other bacteria, they were found to be most closely related to Escherichia coli GyrB (87.6% identity), Klebsiella pneumoniae ParC (78.8% identity), and Salmonella Typhimurium ParE (89.5% identity), respectively. The two topoisomerase genes, parC and parE, were found to be contiguous on the E. tarda chromosome. All 18 quinolone-resistant isolates obtained from Korea thus far did not contain subunit alternations apart from a substitution in GyrA (Ser83$\rightarrow$Arg). However, an alteration in the QRDR of ParC (Ser84$\rightarrow$Ile) following an amino acid substitution in GyrA (Asp87$\rightarrow$Gly) was detected in E. tarda mutants selected in vitro at $8{\mu}g/ml$ ciprofloxacin (CIP). A mutant with a GyrB (Ser464$\rightarrow$Leu) and GyrA (Asp87$\rightarrow$Gly) substitution did not show a significant increase in the minimum inhibitory concentration (MIC) of CIP. None of the in vitro mutants exhibited mutations in parE. Thus, gyrA and parC should be considered to be the primary and secondary targets, respectively, of quinolones in E. tarda.

MicroRNA-214 Regulates the Acquired Resistance to Gefitinib via the PTEN/AKT Pathway in EGFR-mutant Cell Lines

  • Wang, Yong-Sheng;Wang, Yin-Hua;Xia, Hong-Ping;Zhou, Song-Wen;Schmid-Bindert, Gerald;Zhou, Cai-Cun
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.1
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    • pp.255-260
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    • 2012
  • Patients with non-small cell lung cancer (NSCLC) who have activating epidermal growth factor receptor (EGFR) mutations derive clinical benefit from treatment with EGFR-tyrosine kinase inhibitors ((EGFR-TKIs)-namely gefitinib and erlotinib. However, these patients eventually develop resistance to EGFR-TKIs. Despite the fact that this acquired resistance may be the result of a secondary mutation in the EGFR gene, such as T790M or amplification of the MET proto-oncogene, there are other mechanisms which need to be explored. MicroRNAs (miRs) are a class of small non-coding RNAs that play pivotal roles in tumorigenesis, tumor progression and chemo-resistance. In this study, we firstly successfully established a gefitinib resistant cell line-HCC827/GR, by exposing normal HCC827 cells (an NSCLC cell line with a 746E-750A in-frame deletion of EGFR gene) to increasing concentrations of gefitinib. Then, we found that miR-214 was significantly up-regulated in HCC827/GR. We also showed that miR-214 and PTEN were inversely expressed in HCC827/GR. Knockdown of miR-214 altered the expression of PTEN and p-AKT and re-sensitized HCC827/GR to gefitinib. Taken together, miR-214 may regulate the acquired resistance to gefitinib in HCC827 via PTEN/AKT signaling pathway. Suppression of miR-214 may thus reverse the acquired resistance to EGFR-TKIs therapy.

Disulfide Bond as a Structural Determinant of Prion Protein Membrane Insertion

  • Shin, Jae Yoon;Shin, Jae Il;Kim, Jun Seob;Yang, Yoo Soo;Shin, Yeon-Kyun;Kim, Kyeong Kyu;Lee, Sangho;Kweon, Dae-Hyuk
    • Molecules and Cells
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    • v.27 no.6
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    • pp.673-680
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    • 2009
  • Conversion of the normal soluble form of prion protein, PrP ($PrP^C$), to proteinase K-resistant form ($PrP^{Sc}$) is a common molecular etiology of prion diseases. Proteinase K-resistance is attributed to a drastic conformational change from ${\alpha}$-helix to ${\beta}$-sheet and subsequent fibril formation. Compelling evidence suggests that membranes play a role in the conformational conversion of PrP. However, biophysical mechanisms underlying the conformational changes of PrP and membrane binding are still elusive. Recently, we demonstrated that the putative transmembrane domain (TMD; residues 111-135) of Syrian hamster PrP penetrates into the membrane upon the reduction of the conserved disulfide bond of PrP. To understand the mechanism underlying the membrane insertion of the TMD, here we explored changes in conformation and membrane binding abilities of PrP using wild type and cysteine-free mutant. We show that the reduction of the disulfide bond of PrP removes motional restriction of the TMD, which might, in turn, expose the TMD into solvent. The released TMD then penetrates into the membrane. We suggest that the disulfide bond regulates the membrane binding mode of PrP by controlling the motional freedom of the TMD.

Engineering the Cellular Protein Secretory Pathway for Enhancement of Recombinant Tissue Plasminogen Activator Expression in Chinese Hamster Ovary Cells: Effects of CERT and XBP1s Genes

  • Rahimpour, Azam;Vaziri, Behrouz;Moazzami, Reza;Nematollahi, Leila;Barkhordari, Farzaneh;Kokabee, Leila;Adeli, Ahmad;Mahboudi, Fereidoun
    • Journal of Microbiology and Biotechnology
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    • v.23 no.8
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    • pp.1116-1122
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    • 2013
  • Cell line development is the most critical and also the most time-consuming step in the production of recombinant therapeutic proteins. In this regard, a variety of vector and cell engineering strategies have been developed for generating high-producing mammalian cells; however, the cell line engineering approach seems to show various results on different recombinant protein producer cells. In order to improve the secretory capacity of a recombinant tissue plasminogen activator (t-PA)-producing Chinese hamster ovary (CHO) cell line, we developed cell line engineering approaches based on the ceramide transfer protein (CERT) and X-box binding protein 1 (XBP1) genes. For this purpose, CERT S132A, a mutant form of CERT that is resistant to phosphorylation, and XBP1s were overexpressed in a recombinant t-PA-producing CHO cell line. Overexpression of CERT S132A increased the specific productivity of t-PA-producing CHO cells up to 35%. In contrast, the heterologous expression of XBP1s did not affect the t-PA expression rate. Our results suggest that CERT-S132A-based secretion engineering could be an effective strategy for enhancing recombinant t-PA production in CHO cells.

1-Aminocyclopropane-1-Carboxylate Deaminase from Pseudomonas stutzeri A1501 Facilitates the Growth of Rice in the Presence of Salt or Heavy Metals

  • Han, Yunlei;Wang, Rui;Yang, Zhirong;Zhan, Yuhua;Ma, Yao;Ping, Shuzhen;Zhang, Liwen;Lin, Min;Yan, Yongliang
    • Journal of Microbiology and Biotechnology
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    • v.25 no.7
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    • pp.1119-1128
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    • 2015
  • 1-Aminocyclopropane-1-carboxylate (ACC) deaminase, which is encoded by some bacteria, can reduce the amount of ethylene, a root elongation inhibitor, and stimulate the growth of plants under various environmental stresses. The presence of ACC deaminase activity and the regulation of ACC in several rhizospheric bacteria have been reported. The nitrogen-fixing Pseudomonas stutzeri A1501 is capable of endophytic association with rice plants and promotes the growth of rice. However, the functional identification of ACC deaminase has not been performed. In this study, the proposed effect of ACC deaminase in P. stutzeri A1501 was investigated. Genome mining showed that P. stutzeri A1501 carries a single gene encoding ACC deaminase, designated acdS. The acdS mutant was devoid of ACC deaminase activity and was less resistant to NaCl and NiCl2 compared with the wild-type. Furthermore, inactivation of acdS greatly impaired its nitrogenase activity under salt stress conditions. It was also observed that mutation of the acdS gene led to loss of the ability to promote the growth of rice under salt or heavy metal stress. Taken together, this study illustrates the essential role of ACC deaminase, not only in enhancing the salt or heavy metal tolerance of bacteria but also in improving the growth of plants, and provides a theoretical basis for studying the interaction between plant growth-promoting rhizobacteria and plants.

Genotoxicity of Aluminum Oxide ($Al_2O_3$) Nanoparticle in Mammalian Cell Lines

  • Kim, Youn-Jung;Choi, Han-Saem;Song, Mi-Kyung;Youk, Da-Young;Kim, Ji-Hee;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • v.5 no.2
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    • pp.172-178
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    • 2009
  • Nanoparticles are small-scale substances (<100 nm) with unique properties, complex exposure and health risk implications. Aluminum oxide ($Al_2O_3$) nanoparticles (NP) have been widely used as abrasives, wear-resistant coatings on propeller shafts of ships, to increase the specific impulse per weight of composite propellants used in solid rocket fuel and as drug delivery systems to increase solubility. However, recent studies have shown that nano-sized aluminum (10 nm in diameter) can generate adverse effects, such as pulmonary response. The cytotoxicity and genotoxicity of $Al_2O_3$ NP were investigated using the dye exclusion assay, the comet assay, and the mouse lymphoma thymidine kinase (tk$^{+/-}$) gene mutation assay (MLA). IC$_{20}$ values of $Al_2O_3$ NP in BEAS-2B cells were determined the concentration of 273.44 $\mu$g/mL and 390.63 $\mu$g/mL with and without S-9. However IC$_{20}$ values of $Al_2O_3$ NP were found nontoxic in L5178Y cells both of with and without S-9 fraction. In the comet assay, L5178Y cells and BEAS-2B cells were treated with $Al_2O_3$ NP which significantly increased 2-fold tail moment with and without S-9. Also, the mutant frequencies in the $Al_2O_3$ NP treated L5178Y cells were increased compared to the vehicle controls with S-9. The results of this study indicate that $Al_2O_3$ NP can cause primary DNA damage and cytotoxicity but not mutagenicity in cultured mammalian cells.