• 제목/요약/키워드: biological laboratory

검색결과 2,132건 처리시간 0.027초

Scratching Stimuli of Mycelia Influence Fruiting Body Production and ROS-Scavenging Gene Expression of Cordyceps militaris

  • Liu, Gui-Qing;Qiu, Xue-Hong;Cao, Li;Han, Ri-Chou
    • Mycobiology
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    • 제46권4호
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    • pp.382-387
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    • 2018
  • The entomopathogenic fungus Cordyceps militaris is a valuable medicinal ascomycete, which degenerates frequently during subsequent culture. To avoid economic losses during industrialized production, scratching stimuli of mycelia was introduced to improve the fruiting body production. The present results indicated that higher yields and biological efficiency were obtained from two degenerate strains (YN1-14 and YN2-7) but not from g38 (an insertional mutant in Rhf1 gene with higher yields and shorter growth periods). Furthermore, the growth periods of the fruiting bodies were at least 5 days earlier when the mycelia were scratched before stromata differentiation. Three ROS-scavenging genes including Cu/Zn superoxide dismutase (CmSod1), Glutathione peroxidase (CmGpx), and Catalase A (CmCat A) were isolated and their expression profiles against scratching were determined in degenerate strain YN1-14 and mutant strain g38. At day 5 after scratching, the expression level of CmGpx significantly decreased for strain g38, but that of CmSod1 significantly increased for YN1-14. These results indicated that scratching is an effective way to promote fruiting body production of degenerate strain, which may be related at least with Rhf1 and active oxygen scavenging genes.

Radish phospholipid hydroperoxide glutathione peroxidase provides protection against hydroperoxide-mediated injury in mouse 3T3 fibroblasts

  • Li, Tian;Liu, Guan-Lan;Duan, Ming-Xing;Liu, Jin-Yuan
    • BMB Reports
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    • 제42권10호
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    • pp.648-654
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    • 2009
  • Overexpression of phospholipid hydroperoxide glutathione peroxidase (PHGPx) genes has been reported to play an important role in protecting host cells from oxidative injury in several model systems. A radish phospholipid hydroperoxide glutathione peroxidase (RsPHGPx) known to have high catalytic activity was applied to mouse 3T3 fibroblasts to determine the protective effects of PHGPx against oxidative injury triggered by hydroperoxides such as hydrogen peroxide ($H_2O_2$), tert-butyl hydroperoxide (t-BHP) and phosphatidylcholine hydroperoxide (PCOOH). We observed that preincubation of cells with RsPHGPx significantly increased cell viability, reduced levels of malondialdehyde (MDA), inhibited generation of reactive oxygen species (ROS), and maintained natural cell shapes after treatment with $H_2O_2$, t-BHP or PCOOH, indicating that the exogenous RsPHGPx can act as an effective hydroperoxide-scavenger and may also protect target cells from oxidative damage. These results suggest the possibility for use of RsPHGPx as a therapeutic protectant.

Potentiation of Apoptin-Induced Apoptosis by Cecropin B-Like Antibacterial Peptide ABPs1 in Human HeLa Cervical Cancer Cell Lines is Associated with Membrane Pore Formation and Caspase-3 Activation

  • Birame, Basse Mame;Wang, Jigui;Yu, Fuxian;Sun, Jiazeng;Li, Zhili;Liu, Weiquan
    • Journal of Microbiology and Biotechnology
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    • 제24권6호
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    • pp.756-764
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    • 2014
  • Apoptin, a chicken anemia virus-encoded protein, induces apoptosis in chicken or human tumor cells, localizing in their nuclei as opposed to the cytoplasm of non-transformed cells. The present study was undertaken to investigate whether ABPs1 could potentiate apoptin-induced apoptosis in HeLa cells. ABPs1 and the apoptin genes were successfully cloned into pIRES2-EGFP expression vector and expressed in HeLa cells. We report that ABPs1 augments apoptin cell growth inhibition in a concentration- and time-dependent manner. The DAPI staining and scanning electron microscopy observations revealed apoptotic bodies and plasma membrane pores, which were attributed to apoptin and ABPs1, respectively. Further, ABPs1 in combination with apoptin was found to increase the expression of Bax and to decrease the expression of survivin compared with either agent alone or the control. The apoptotic rate of HeLa cells treated with ABPs1 and apoptin in combination for 48 h was 53.95%. The two-gene combination increased the caspase-3 activity of HeLa cells. Taken together, our study suggests that ABPs1 combined with apoptin significantly inhibits HeLa cell proliferation, and induces cell apoptosis through membrane defects, up-regulation of Bax expression, down-regulation of survivin expression, and activation of the caspase-3 pathway. Thus, the combination of ABPs1 and apoptin could serve as a means to develop novel gene therapeutic agents against human cervical cancer.

Characterization of Two GAS1 Genes and Their Effects on Expression and Secretion of Heterologous Protein Xylanase B in Kluyveromyces lactis

  • Lian, Zhao;Jiang, Jing-Bo;Chi, Shuang;Guan, Guo-Hua;Li, Ying;Li, Ji-Lun
    • Journal of Microbiology and Biotechnology
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    • 제25권12호
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    • pp.1977-1988
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    • 2015
  • β-1,3-glucanosyltransferases play essential roles in cell wall biosynthesis in yeast. Kluyveromyces lactis has six putative β-1,3-glucanosyltransferase genes. KlGAS1-1 and KlGAS1-2 are homologs of Saccharomyces cerevisiae gene GAS1. RT-qPCR indicated the transcription level of KlGAS1-1 was significantly reduced while heterologous protein (thermostable xylanase B) secretion was enhanced during medium optimization. To evaluate if these two events were related, and to improve xylanase B secretion in K. lactis, we constructed KlGAS1-1 and KlGAS1-2 single deletion strains and double deletion strain, respectively. KlGAS1-1 gene deletion resulted in the highest xylanase B activity among the three mutants. Only the double deletion strain showed morphology similar to that of the GAS1 deletion mutant in S. cerevisiae. The two single deletion strains differed in terms of cell wall thickness and xylanase B secretion. Transcription levels of β-1,3-glucanosyltransferase genes and genes related to protein secretion and transport were assayed. The β-1,3-glucanosyltransferase genes displayed transcription complementation in the cell wall synthesis process. KlGAS1-1 and KlGAS1-2 affected transcription levels of secretion- and transport-related genes. Differences in protein secretion ratio among the three deletion strains were associated with changes of transcription levels of secretion- and transport-related genes. Our findings indicate that KlGAS1-1 deletion is an effective tool for enhancing industrial-scale heterologous protein secretion in K. lactis.

Improvement of Natamycin Production by Cholesterol Oxidase Overexpression in Streptomyces gilvosporeus

  • Wang, Miao;Wang, Shaohua;Zong, Gongli;Hou, Zhongwen;Liu, Fei;Liao, D. Joshua;Zhu, Xiqiang
    • Journal of Microbiology and Biotechnology
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    • 제26권2호
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    • pp.241-247
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    • 2016
  • Natamycin is a widely used antifungal antibiotic. For natamycin biosynthesis, the gene pimE encodes cholesterol oxidase, which acts as a signalling protein. To confirm the positive effect of the gene pimE on natamycin biosynthesis, an additional copy of the gene pimE was inserted into the genome of Streptomyces gilvosporeus 712 under the control of the ermE* promoter (permE*) using intergeneric conjugation. Overexpression of the target protein engendered 72% and 81% increases in the natamycin production and cell productivity, respectively, compared with the control strain. Further improvement in the antibiotic production was achieved in a 1 L fermenter to 7.0 g/l, which was a 153% improvement after 120 h cultivation. Exconjugants highly expressing pimE and pimM were constructed to investigate the effects of both genes on the increase of natamycin production. However, the co-effect of pimE and pimM did not enhance the antibiotic production obviously, compared with the exconjugants highly expressing pimE only. These results suggest not only a new application of cholesterol oxidase but also a useful strategy to genetically engineer natamycin production.

Impairment of a parabolic bursting rhythm by the ectopic expression of a small conductance $Ca^{2+}$-activated $K^+$ channel in Aplysia neuron R15

  • Lee, Yong;Han, Jin-Hee;Lim, Chae-Seok;Chang, Deok-Jin;Lee, Yong-Seok;Heun Soh;Park, Chul-Seung;Kaang, Bong-Kiun
    • 한국생물물리학회:학술대회논문집
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    • 한국생물물리학회 2003년도 정기총회 및 학술발표회
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    • pp.38-38
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    • 2003
  • The electrical properties of neurons are produced by the coordinated activity of ion channels (Hille, 1992). $K^{+}$ channels play a key role in shaping action potentials and in determining neural firing patterns. Small conductance $Ca^{2+}$-activated $K^{+}$ (S $K_{Ca}$ ) channels are involved in modulating the slow component of afterhyperpolarization (AHP) (Kohler et al., 1996). Here we examine whether rat type 2 S $K_{Ca}$ (rSK2) channels can affect the shape of the action potential and the neural firing pattern, by overexpressing rat SK2 channels in Aplysia neuron R15. Our results show that rSK2 overexpression decreased the intraburst frequency and changed the regular bursting activity of neurons to an irregular bursting or beating pattern in R15, Furthermore, the overexpression of rSK2 channels increased AHP and reduced the duration of the action potential. Thus, our results suggest that ectopic S $K_{Ca}$ channels play an important role in regulating the filing pattern and the shape of the action potential.ntial.

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Recent Progresses in the Linguistic Modeling of Biological Sequences Based on Formal Language Theory

  • Park, Hyun-Seok;Galbadrakh, Bulgan;Kim, Young-Mi
    • Genomics & Informatics
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    • 제9권1호
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    • pp.5-11
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    • 2011
  • Treating genomes just as languages raises the possibility of producing concise generalizations about information in biological sequences. Grammars used in this way would constitute a model of underlying biological processes or structures, and that grammars may, in fact, serve as an appropriate tool for theory formation. The increasing number of biological sequences that have been yielded further highlights a growing need for developing grammatical systems in bioinformatics. The intent of this review is therefore to list some bibliographic references regarding the recent progresses in the field of grammatical modeling of biological sequences. This review will also contain some sections to briefly introduce basic knowledge about formal language theory, such as the Chomsky hierarchy, for non-experts in computational linguistics, and to provide some helpful pointers to start a deeper investigation into this field.