• Title/Summary/Keyword: wild type

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Isolation and characterization of Vitreoscilla mutant defective in catalase-peroxidase hydroperoxidase I

  • Kim, Hee-Jung;Moon, Ja-Young;Lee, John-Hwa;Park, Kie-In
    • Korean Journal of Veterinary Service
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    • v.30 no.3
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    • pp.291-304
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    • 2007
  • Mutants of an obligate aerobic bacterium, Vitreoscilla, that have deficiency in heat-labile catalase-peroxidase hydroperoxidase I (HPI) were created by EMS treatment. The catalase-peroxidase HPI-deficient mutant showed substantially lower peroxidase activity in exponential and mid-stationary phase compared with the wild type strain. In late stationary phase, the mutant exhibited no peroxidase activity. Peroxidase deficiency in the mutant was revealed by polyacrylamide gels stained for peroxidase activity. Characteristically, catalase levels in the mutant increased about 14- and 8-fold during growth in the exponential and stationary phases, respectively, compared to those in the wild type, suggesting a compensatory effect for protection from $H_2O_2$ toxicity. The mutant showed differences in physiology from the wild type: retardation in growth rate and decrease in oxygen consumption. Both the wild type and the catalase-peroxidase HPI-deficient mutant of Vitreoscilla had lower growth rates in media containing increasing $H_2O_2$ concentrations. However, the mutant exhibited an additionally decreased growth rate after 6 to 8 h of growth compared to the wild type. The wild type was resistent up to 20 mM $H_2O_2$, whereas the mutant was very sensitive to high concentrations of exogenous $H_2O_2$. Although elevated catalase levels would provide protection of the bacteria from the deleterious effect of $H_2O_2$, it did not appear to be complete. Cell-free extracts of the mutant showed decreased NADH oxidation rates and higher accumulation of $H_2O_2$ during this oxidation. These results may account for the impaired growth and earlier onset of death phase by the catalase-peroxidase HPI-deficient mutant of Vitreoscilla.

Studies on the Escherichia coli Hemolysin Antigenic Sites and Functional Sites for the Hemolysin Vaccine Development (Hemolysin 백신 개발을 위한 요로계 감염 대장균들의 Hemolysin Antigenic Sites, Functional Sites 상동성 연구)

  • 지근억;백광현
    • Microbiology and Biotechnology Letters
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    • v.20 no.3
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    • pp.301-310
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    • 1992
  • This work was performed to investigate the possibility of using J96 hemolysin(Hly, Hly A) vaccine against urinary tract infecting Escherichia coli. Based on the known sequence of J96 hemolysin which was originally isolated from a pyelonephritis patient, ten 20-mer oligonucleotide probes were synthesized. Radioactive labelled 8 probes showed positive colony blots against most of the hemolysin producing wild type E. coli, while HA484 and HA661 showed 28.3, 71.7% positive blots, respectively. This result means that hemolysin genes are highly conserved. Also, 12 anti-Hly MABs(monoclonal antibodies) showed more than 90% positive immunoblots against secreted hemolysin from wild type E. coli. Especially, the result that MAB132 neutralized hemolysin from all of the wild type E. coli augments the idea that hemolysin will be effective as a vaccine.

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Increased Caveolin-2 Expression in Brain Endothelial Cells Promotes Age-Related Neuroinflammation

  • Hyunju, Park;Jung A, Shin;Jiwoo, Lim;Seulgi, Lee;Jung-Hyuck, Ahn;Jihee Lee, Kang;Youn-Hee, Choi
    • Molecules and Cells
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    • v.45 no.12
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    • pp.950-962
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    • 2022
  • Aging is a major risk factor for common neurodegenerative diseases. Although multiple molecular, cellular, structural, and functional changes occur in the brain during aging, the involvement of caveolin-2 (Cav-2) in brain ageing remains unknown. We investigated Cav-2 expression in brains of aged mice and its effects on endothelial cells. The human umbilical vein endothelial cells (HUVECs) showed decreased THP-1 adhesion and infiltration when treated with Cav-2 siRNA compared to control siRNA. In contrast, Cav-2 overexpression increased THP-1 adhesion and infiltration in HUVECs. Increased expression of Cav-2 and iba-1 was observed in brains of old mice. Moreover, there were fewer iba-1-positive cells in the brains of aged Cav-2 knockout (KO) mice than of wild-type aged mice. The levels of several chemokines were higher in brains of aged wild-type mice than in young wild-type mice; moreover, chemokine levels were significantly lower in brains of young mice as well as aged Cav-2 KO mice than in their wild-type counterparts. Expression of PECAM1 and VE-cadherin proteins increased in brains of old wild-type mice but was barely detected in brains of young wild-type and Cav-2 KO mice. Collectively, our results suggest that Cav-2 expression increases in the endothelial cells of aged brain, and promotes leukocyte infiltration and age-associated neuroinflammation.

Bird Species Diversity Analysis According to the Type of Forest Vegetation (산림식생유형에 따른 조류 종다양성 분석)

  • Park, In-Hwan;Kim, Yu-Hoon;Cho, Kwang-Jin
    • Journal of the Korean Society of Environmental Restoration Technology
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    • v.15 no.6
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    • pp.43-52
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    • 2012
  • For this paper, a study targeting 9 forest patches in Dangjin to suggest for the wild birds, was conducted. The correlation between the emergence of wild birds and the forest vegetation was analyzed, and the patterns of forest vegetation for the improvement of species diversity were suggested by classifying wild birds and foraging guild by the type of forest vegetation. As for the characteristics of the wild birds emerged, more species and bigger population appeared in the vegetation type of mixed stand forest where the natural deciduous broadleaf trees and evergreen needleleaf trees coexisted in the large scale vegetation area. Thus, it was known that the advent of summer migratory birds and resident birds were affected the most by forest patches and vegetation types. As for foraging guild, the larger the forest paches the more the species and the population of the wild birds inhabiting in shrubs and bushes.

Strain Improvement of Candida tropicalis for the Production of Xylitol: Biochemical and Physiological Characterization of Wild-type and Mutant Strain CT-OMV5

  • Rao Ravella Sreenivas;Jyothi Cherukuri Pavanna;Prakasham Reddy Shetty;Rao Chaganti Subba;Sarma Ponnupalli Nageshwara;Rao Linga Venkateswar
    • Journal of Microbiology
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    • v.44 no.1
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    • pp.113-120
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    • 2006
  • Candida tropicalis was treated with ultraviolet (UV) rays, and the mutants obtained were screened for xylitol production. One of the mutants, the UV1 produced 0.81g of xylitol per gram of xylose. This was further mutated with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), and the mutants obtained were screened for xylitol production. One of the mutants (CT-OMV5) produced 0.85g/g of xylitol from xylose. Xylitol production improved to 0.87 g/g of xylose with this strain when the production medium was supplemented with urea. The CT-OMV5 mutant strain differs by 12 tests when compared to the wild-type Candida tropicalis strain. The XR activity was higher in mutant CT-OMV5. The distinct difference between the mutant and wild-type strain is the presence of numerous chlamvdospores in the mutant. In this investigation, we have demonstrated that mutagenesis was successful in generating a superior xylitol-producing strain, CT-OMV5, and uncovered distinctive biochemical and physiological characteristics of the wild-type and mutant strain, CT-OMV5.

Growth, quality, and yield characteristics of transgenic potato (Solanum tuberosum L.) overexpressing StMyb1R-1 under water deficit

  • Im, Ju-Sung;Cho, Kwang-Soo;Cho, Ji-Hong;Park, Young-Eun;Cheun, Chung-Gi;Kim, Hyun-Jun;Cho, Hyun-Mook;Lee, Jong-Nam;Jin, Yong-Ik;Byun, Myung-Ok;Kim, Dool-Yi;Kim, Myeong-Jun
    • Journal of Plant Biotechnology
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    • v.39 no.3
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    • pp.154-162
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    • 2012
  • This study was conducted to evaluate agronomic characteristics such as growth, quality, and yields of StMyb1R-1 transgenic potato and also to obtain the basic data for establishing assessment guidelines of transgenic potato. Three transgenic lines (Myb 1, Myb 2, and Myb 8) were cultivated under conventional irrigation, drought condition, and severe drought condition and were analyzed by comparing with wild type, non-transgenic cv. Superior. Myb 2 showed a different flower color from wild type and Myb 1 had much bigger secondary leaflets than wild type. Myb 1 and Myb 2 showed higher $P_2O_5$ content in both top and root zone and longer shaped tubers than wild type. In yield factors, transgenic lines had more tubers than wild type, however their yield decreases were severe because of the poor enlargement of tuber under water deficit condition. This tendency was noticeable in Myb 1 and Myb 2. In TR ratio, chlorophyll content, dry matter rate, and relative water content, there were no big differences between transgenic lines and wild type. Meanwhile, in phenotype, growth, quality, and yield factors, substantial equivalent was confirmed between Myb 8 and wild type. Then, Myb 8 showed the highest marketable tuber yield under conventional irrigation, while showed lower level than wild type under water deficit. Judged by this result, the enhancing droughttolerance by StMyb1R-1 gene might actually not mean the enhancement of photosynthesis or starch accumulation in tuber and, furthermore, not the yield improvement. More detailed research will be required to accurately understand the relationship between StMyb1R-1 and yield factors.

Evaluation of Nitrogen Fixation between Supernodulating Soybean Mutants and their Wild-Types Using $^{15}N$ in Field Conditions

  • Youn, Jong-Tag;Van, Kyu-Jung;Ryu, Yong-Hwan;Shimada, Shinji;Lee, Choon-Ki;Kim, Sun-Lim;Seo, Sea-Jung;Lee, Suk-Ha
    • Journal of Crop Science and Biotechnology
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    • v.11 no.1
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    • pp.31-38
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    • 2008
  • Improvement of $N_2$ fixation by symbiotic rhizobia is considered an effective means for enhancing its productivity without high input of nitrogen(N) fertilizer. Several methods to improve $N_2$ fixation have been proposed including the use of supernodulating mutants. The objective of this research was to identify the varietal difference in N and $N_2$-fixation ability among the soybean supernodulating mutants, SS2-2 and Sakukei 4, with different nodulation abilities using $^{15}N$ in field conditions in both Korea and Japan. The dry weight(DW) was higher in wild-type soybeans. The distribution rate of DW in each plant part was high in seeds of supernoduating and wild-type soybeans but high in stems and leaves of non-nodulating mutants. Although the supernodulating mutants had a low DW rate at maturity, they showed a similar $N_2$ fixation ability compared with wild-type. Supernodulating mutant plants mainly obtained N from $N_2$ fixation, while soil N was the main resource for obtaining N in non-nodulating mutants. The percentage of N derived from atmospheric dinitrogen(Ndfa) was higher in supernodulating mutants than in wild-type and relatively high in seeds between plant parts at maturity. In particular, supernodulating mutants showed higher N content in roots than those of wild-type and non-nodulating mutants. It was considered that supernodulating mutants have the advantage of saving nitrate in soil and being beneficial for N absorption of subsequent crops due to their conserving more N in the field and releasing considerable amounts of N from roots and leaves fallen to the soil.

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A TaqMan Real-Time PCR Assay for Quantifying Type III Hepatopancreatic Parvovirus Infections in Wild Broodstocks and Hatchery-Reared Postlarvae of Fenneropenaeus chinensis in Korea

  • Jang, In-Kwon;Suriakala, Kannan;Kim, Jong-Sheek;Meng, Xian-Hong;Choi, Tae-Jin
    • Journal of Microbiology and Biotechnology
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    • v.21 no.11
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    • pp.1109-1115
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    • 2011
  • A highly sensitive and specific TaqMan real-time PCR was used to quantify hepatopancreatic parvovirus (HPV) type III infections in wild broodstocks and hatchery-reared postlarvae (PL) of Fenneropenaeus chinensis. Totals of 159 and 162 wild brooders from three locations were captured, and 140 and 180 PL were obtained from seven and six commercial hatcheries in 2007 and 2008, respectively. Among the three wild broodstock groups from 2007, only 1 group showed HPV infection and 3.2% of 159 brooders were positive for HPV infection. In 2008, HPV infections were observed from all three wild broodstock groups with $1.93{\times}10^4$ copies/mg tissue of pleopods. Of 162 brooders, 26.6% were positive for HPV infection. No PL from the two hatcheries collected in 2007 showed HPV infection, and PL from the rest of the five hatcheries had up to $1.74{\times}10^6$ copies/ng of DNA, and PL from three hatcheries showed HPV infections with over 1,000 copies/ng of DNA. The PL from all seven hatcheries collected in 2008 showed up to $2.10{\times}10^5$ HPV copies/ng of DNA. PL from two hatcheries showed less than 100 copies/ng of DNA, but PL from the rest of the hatcheries showed HPV infections with over 1,000 copies/ng of DNA. These results show that HPV type III is widely distributed in Korea in addition to previously reported HPV type I, and they can be effectively detected by type-specific realtime PCR.

Influence of Respiration on Photosynthetic Electron Transport in psaB Mutants from Cyanobacterium Synechocystis sp. PCC6803 (Cyanobacterium Synechocystis sp.PCC6803 psaB 돌연변이주의 광합성 전자전달에 미치는 호흡의 영향)

  • 윤병철;장남기
    • Asian Journal of Turfgrass Science
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    • v.11 no.1
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    • pp.59-72
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    • 1997
  • The influence of respiration on photosythetic electron transport were investigated in the Wid type and psaB mutants from Syneehocystis sp. PCC6803. The amount of glucose uptake in the wild type was proportional to the glucose concentration added in wild type and less than that of psaB mutants in the dark. It was suggested that psaB mutants more depend on the glucose than the wild type. It was investigated how the activities of isocitrate dehydrogenase(IDH) and glucose-6-phos-phate dehydrogenase(G6PDH) were changed. The activities of IDH were very low. While, the ac-tivities of G6PDH were much higher than that of IDH. These results agree to the reports that ex-ogenous glucose was dismilated aerobically via Oxidative Pentose Phosphate Pathway in heterotrophic cyanobacteria. PsaB mutants showed high G6PDH activity in the presence of glucose as well as in the dark and high respiratory activities especially in the dark. It was also investigated how photosynthetic electron transport activities were changed. PsaB mutants showed higher photosynthetic electron tranasport activities than wild type in the presence of glucose as well as in the dark. In the results, it was proposed that photosynthetic electron transport between PS I and PS U was complemented by respiratory electron transport through the NADPH generated by Dxidative Pentose Phophate Pathway in psaB mutant from Synechocystis sp. PCC6803. Key words: Photosynthetic electron transport, Respiration, Synechoystis sp. PCC6803, psaB mutant, Glucose uptake, IDH, G6PDH, Respiratory electron transport activity.

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Characterization of Chlorella Vulgaris Mutants Generated by EMS (Ethyl Methane Sulphonate) (EMS (Ethyl Methane Sulphonate) 처리에 의한 Chlorella Vulgaris 변이주 생성 및 특성 분석)

  • Kim, Ok Ju;Lee, Jae-Hwa
    • Applied Chemistry for Engineering
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    • v.26 no.3
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    • pp.265-269
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    • 2015
  • Chlorella vulgaris (C. vulgaris) is a spherical unicellular green algae and the diameter ranges from 2 to $10{\mu}m$. C. vulgaris possess nutritional excellence because it contains various functional materials including high protein contents, chlorophyll, carotenoid, and chlorella growth factor (CGF). In order to study effects of mutagen, ethyl methane sulphonate (EMS) was used as a chemical mutagen and some mutants could be obtained. We named 2 type mutants as E14 and E24 obtained after treating with EMS. In the cell growth, growth patterns of mutants were similar to those of the wild type. Chlorophyll contents of E14 and E24 increased up to 99 and 52%, respectively compared to those of the wild type. The carotenoid content of E14 increased to 7%, but the value of E24 decreased 5% compared to that of the wild type. For the lipid contents E24 increased to 23%, while E14 decreased 12% when compared to those of the wild type. As a result, there is no difference between the mutants and wild type in the cell growth, but considering that mutants contains more physiological materials than those of the wild type, we can expect the mutants of C. vulgaris could be used as important high added-value materials.