• Title/Summary/Keyword: Pigment analyses

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Functional Characterization of Genes Located at the Aurofusarin Biosynthesis Gene Cluster in Gibberella zeae

  • Kim, Jung-Eun;Kim, Jin-Cheol;Jin, Jian-Ming;Yun, Sung-Hwan;Lee, Yin-Won
    • The Plant Pathology Journal
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    • v.24 no.1
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    • pp.8-16
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    • 2008
  • Aurofusarin is a polyketide pigment produced by some Fusarium species. The PKS12 and GIP1 genes, which encode a putative type I polyketide synthase (PKS) and a fungal laccase, respectively, are known to be required for aurofusarin biosynthesis in Gibberella zeae (anamorph: Fusarium graminearum). The ten additional genes, which are located within a 30 kb region of PKS12 and GIP1 and regulated by a putative transcription factor (GIP2), organize the aurofusarin biosynthetic cluster. To determine if they are essential for aurofusarin production in G. zeae, we have employed targeted gene deletion, complementation, and chemical analyses. GIP7, which encodes O-methyltransferase, is confirmed to be required for the conversion of norrubrofusarin to rubrofusarin, an intermediate of aurofusarin. GIP1-, GIP3-, and GIP8-deleted strains accumulated rubrofusarin, indicating those gene products are essential enzymes for the conversion of rubrofusarin to aurofusarin. Based on the phenotypic changes in the gene deletion strains examined, we propose a possible pathway for aurofusarin biosynthesis in G. zeae. Our results would provide important information for better understanding of naphthoquinone biosynthesis in other fdarnentous fungi as well as the aurofusarin biosynthesis in G. zeae.

Innate Color Preference of Zebrafish and Its Use in Behavioral Analyses

  • Park, Jong-Su;Ryu, Jae-Ho;Choi, Tae-Ik;Bae, Young-Ki;Lee, Suman;Kang, Hae Jin;Kim, Cheol-Hee
    • Molecules and Cells
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    • v.39 no.10
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    • pp.750-755
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    • 2016
  • Although innate color preference of motile organisms may provide clues to behavioral biases, it has remained a longstanding question. In this study, we investigated innate color preference of zebrafish larvae. A cross maze with different color sleeves around each arm was used for the color preference test (R; red, G; green, B; blue, Y; yellow). The findings showed that 5 dpf zebrafish larvae preferred blue over other colors (B > R > G > Y). To study innate color recognition further, tyrosinase mutants were generated using CRISPR/Cas9 system. As a model for oculocutaneous albinism (OCA) and color vision impairment, tyrosinase mutants demonstrated diminished color sensation, indicated mainly by hypopigmentation of the retinal pigment epithelium (RPE). Due to its relative simplicity and ease, color preference screening using zebrafish larvae is suitable for high-throughput screening applications. This system may potentially be applied to the analysis of drug effects on larval behavior or the detection of sensory deficits in neurological disorder models, such as autism-related disorders, using mutant larvae generated by the CRISPR/Cas9 technique.

A Novel Rapid Fungal Promoter Analysis System Using the Phosphopantetheinyl Transferase Gene, npgA, in Aspergillus nidulans

  • Song, Ha-Yeon;Choi, Dahye;Han, Dong-Min;Kim, Dae-Hyuk;Kim, Jung-Mi
    • Mycobiology
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    • v.46 no.4
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    • pp.429-439
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    • 2018
  • To develop a convenient promoter analysis system for fungi, a null-pigment mutant (NPG) of Aspergillus nidulans was used with the 4'-phosphopantetheinyl transferase (PPTase) gene, npgA, which restores the normal pigmentation in A. nidulans, as a new reporter gene. The functional organization of serially deleted promoter regions of the A. nidulans trpC gene and the Cryphonectria parasitica crp gene in filamentous fungi was representatively investigated to establish a novel fungal promoter assay system that depends on color complementation of the NPG mutant with the PPTase npgA gene. Several promoter regions of the trpC and crp genes were fused to the npgA gene containing the 1,034-bp open reading frame and the 966-bp 3' downstream region from the TAA, and the constructed fusions were introduced into the NPG mutant in A. nidulans to evaluate color recovery due to the transcriptional activity of the sequence elements. Serial deletion of the trpC and crp promoter regions in this PPTase reporter assay system reaffirmed results in previous reports by using the fungal transformation step without a laborious verification process. This approach suggests a more rapid and convenient system than conventional analyses for fungal gene expression studies.

Investigation of Wood Species and Conservation Status of Wooden Seated Amitabha Buddha Triad and Wooden Amitabha Buddha Altarpiece of Yongmunsa Temple, Yecheon, Korea (Treasure)

  • CHOI, Jaewan;PARK, Junghae;KIM, Soochul
    • Journal of the Korean Wood Science and Technology
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    • v.50 no.3
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    • pp.193-217
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    • 2022
  • The Wooden Seated Amitabha Buddha Triad and Altarpiece have not been the subjects of definitive species identification and scientific analysis. In this study, visual investigation, portable X-ray fluorescence (p-XRF), species identification, and lacquer layer observations were carried out to determine the original materials and conservation status. Cracks, exfoliation and discoloration were detected during the visual investigation. The p-XRF data indicated that chrome oxide green, malachite, gold, cadmium red, cinnabar, minium, hematite, lead white, ink stick (Carbon), and copper were used for pigments and the coronet accessory. Tilia spp., Abies spp., and Pinus spp. were identified from both the Buddha Triad and Altarpiece. Finally, lacquer layer analyses of the base layer, lacquer layer, orange pigment layer, and gold leaf layer observed modern synthetic pigments likely used in previous conservation. As the Korean Cultural Heritage Charter and the International Charter for the Conservation and Restoration of Monuments and Sites clearly state that objects of cultural heritage must be conserved using their original materials, future conservation of these objects should utilize the data obtained in this study to employ traditional materials. Furthermore, a deterioration map diagnosis can be applied together with the obtained analysis data to understand the conservation status of and inform an appropriate and authentic conservation treatment for the Buddha Triad and Altarpiece.

PREDICTION OF PHYSICO-CHEMICAL AND TEXTURE CHARACTERISTICS OF BEEF BY NEAR INFRARED TRANSMITTANCE SPECTROSCOPY

  • Olivan, Mamen;Delaroza, Begona;Mocha, Mercedes;Martinez, Maria Jesus
    • Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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    • 2001.06a
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    • pp.1256-1256
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    • 2001
  • The physico-chemical and texture characteristics of meat determine the nutritional, technological and sensory quality. However, the analysis of meat quality requires expensive, laborious and time consuming analytical methods. The objective of this study was to evaluate NIR spectroscopy using transmittance for determining the moisture, fat, protein and total pigment content, the water holding capacity (WHC) and the toughness of beef meat. A total of 318 spectra were recorded from ground beef samples by a Feed Analyzer 1265 of Infratec. The samples were obtained from the Longissimus muscle of the 10$^{th}$ rib of yearling bulls, ground with an electrical chopper, vacuum packaged, aged during 7 days and frozen at -24$^{\circ}C$ until the analyses were done. Moisture content was measured by oven drying at 10$0^{\circ}C$, fat content was determined by Soxhlet extraction and protein content was estimated from nitrogen content using the Kjeldahl analysis. The total pigment content was determined by the method of Hornsey and the WHC using the method of filter paper press. The instrumental evaluation of texture (maximum load WB, maximum stress MS and toughness) was conducted in an Instron equipment with a Warner-Bratzler shearing device. This analysis was performed on a chop of 3.5 cm obtained from the longissimus of the 8$^{th}$ rib, aged during 7 days, kept frozen at -24$^{\circ}C$ and cooked before the analysis. Near infrared spectra were recorded as log 1/T (T=transmittance) at 2 nm intervals from 850 to 1050 nm using a Feed Analyzer 1265 of Infratec. Calibrations were performed with the WinISI software (vs. 1.02) using the MPLS method. To examine the effect of scatter correction o. derivation of spectra on the calibration performance, calibrations were calculated with the crude spectra or pretreated with different mathematical treatments (inverse MSC, SNVD) and/or second derivative operation. For chemical composition, the use of the scatter corrections improved the calibration statistics, in terms of lower SECV and higher $r^2$. In most of the variables, the use of the 2$^{nd}$ derivative improved the predictions, mainly when combined with the SNVD treatment. However, for predicting the texture traits, the best estimation was obtained from the crude spectrum. These results showed that the equations obtained for predicting moisture, fat and total pigments were very accurate, with $r^2$ being higher that 0.9. However, the prediction of the texture traits (WB, MS, toughness) from ground meat was poor.

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Partial Characterization of Bacillus thuringiensis var. kurstaki Temperature-sensitive Mutants (Bacillus thringiensis var. kurstaki 감온성 돌연변이주의 일부특성)

  • 김영권;유관희;이형환;이호원
    • Korean Journal of Microbiology
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    • v.23 no.3
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    • pp.203-208
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    • 1985
  • Partial characterization of B. thuringiensis var. kurstaki 3ab temperature-sensitive mutants was carried out through biochemical analyses, utilization tests of carbohydrate sources, antobiotic resistant test, hemolytic reaction test, growth measurement of Fructus gardenia sxtrant medium and toxicity test against mice. Six ts mutants, ts-U154, ts-U601, ts-U602, ts-U603, tsU-604, and ts-U788 could not produce urease, ts-U603 lost its motility, ts-U154 could not use salicin and cellobiose and ts-U603 not ribose. All ts mutants except ts-U154 and wild type strain were resistant to cephalothin, ampicillin, and penicillin. but ts-U154 was sensitive to the three. Four mutants, ts-U21, ts-U74, ts0U131 and ts-U154 did not form pigment colonies on the F. gardenia medium. All the mutants and wild type strain showed hemolysis reaction on the blood agar. The B. thuringiensis and mutants were not toxic to mice.

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Insertional mutations exhibiting high cell-culture density HCD phenotypes are enriched through continuous subcultures in Chlamydomonas reinhardtii

  • Thung, Leena;He, Jing;Zhu, Qingling;Xu, Zhenyu;Liu, Jianhua;Chow, Yvonne
    • ALGAE
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    • v.33 no.1
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    • pp.127-141
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    • 2018
  • Low efficiency in microalgal biomass production was largely attributed to the low density of algal cell cultures. Though mutations that reduced the level of chlorophyll or pigment content increased efficiency of photon usage and thus the cell-culture density under high-illumination growth conditions (e.g., >$500{\mu}mol\;photon\;m^{-2}\;s^{-1}$), it was unclear whether algae could increase cell-culture density under low-illumination conditions (e.g., ${\sim}50{\mu}mol\;photon\;m^{-2}\;s^{-1}$). To address this question, we performed forward genetic screening in Chlamydomonas reinhardtii. A pool of >1,000 insertional mutants was constructed and subjected to continuous subcultures in shaking flasks under low-illumination conditions. Complexity of restriction fragment length polymorphism (RFLP) pattern in cultures indicated the degree of heterogeneity of mutant populations. We showed that the levels of RFLP complexity decreased when cycles of subculture increased, suggesting that cultures were gradually populated by high cell-culture density (HCD) strains. Analysis of the 3 isolated HCD mutants after 30 cycles of subcultures confirmed that their maximal biomass production was 50-100% higher than that of wild type under low-illumination. Furthermore, levels of chlorophyll content in HCD mutant strains were similar to that of wild type. Inverse polymerase chain reaction analysis identified the locus of insertion in two of three HCD strains. Molecular and transcriptomic analyses suggested that two HCD mutants were a result of the gain-of-function phenotype, both linking to the abnormality of mitochondrial functions. Taken together, our results demonstrate that HCD strains can be obtained through continuous subcultures under low illumination conditions.

A Study on Comparison of Phycocyanin Extraction Methods for Hyperspectral Remote Sensing of Cyanobacteria in Turbid Inland Waters (국내 담수역 남조류 원격탐사를 위한 피코시아닌 추출법 비교 연구)

  • Ha, Rim;Shin, Hyunjoo;Nam, Gibeom;Park, Sanghyun;Kang, Taegu;Song, Hyunoh;Lee, Hyuk
    • Journal of Korean Society on Water Environment
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    • v.32 no.6
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    • pp.520-527
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    • 2016
  • Phycocyanin (PC) is one of the water-soluble accessory pigments of cyanobacteria species, and its concentration is used to estimate the presence and relative abundance of cyanobacteria. In laboratory experiments, PC content of field data were determined using Sarada's freeze-thaw method in algal bloom season. The effectiveness of three selected extraction methods (repeated freeze-thaw method, homogenization, power control) for PC were determined. The extraction efficiency of phycocyanin was the highest (of the methods compared) when a single freezing-thawing cycle was followed by pre-sonication. Applying this optimized method to surface water of Korean inland waters, the average concentration distribution was estimated at $2.9{\sim}51.9mg/m^3$. It has been shown that the optimized pre-sonication method is suitable to measure cyanobacteria PC content for the characterization of inland waters. The approach and results of this study indicates the potential of effective methods for remote monitoring and management of water quality in turbid inland waters using hyperspectral remote sensing.

Isolation and Identification of Streptomyces sp. Producing Anti-vancomycin Resistant Staphylococcus aureus Substance (반코마이신 내성 Staphylococcus aureus 억제 물질 생산 Streptomyces sp.의 분리 및 동정)

  • Oh Se-Teak;Lee Jun-Jae;Lee Ji-Youn;Kim Jin-Kyu;Yang Si-Yong;Kim Yang-Soo;Song Min-Dong
    • Microbiology and Biotechnology Letters
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    • v.33 no.2
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    • pp.90-95
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    • 2005
  • An Actinomycetes producing an anti-VRSA (vancomycin-resistant Staphylococcus aureus) substance was isolated from soil. The cultural, morphological, physiological and phylogenetic analyses of an isolated strain were investigated for identification. Cultural characteristics based on ISP (International Streptomyces Project) were as follows: white aerial mycelium, yellow reverse side, and good growth on various medium. Also, the isolate did not produce the soluble pigment. Morphological characteristics were showed cylindrical spore chain and smooth spore surface by SEM (Scanning Electron Microscope). Physiological characteristics were showed LL-type by DAP isomer analysis and detected glycine, glutamic acid and alanine. A phylogenetic analysis of the 16S rDNA provided a clue that the isolated strain was actually a member of the genus Streptomyces, because the determined sequence exhibited a higher homology with Streptomyces echinatus. The isolate was identified to be a genus of Streptomyces sp.. The optimal culture conditions for the maximum production of anti-VRSA substance by Streptomyces sp. were attained in a culture medium composed of $2.0\%$ (w/v) glucose, and $0.4\%$ (w/v) yeast extract. The anti-VRSA substance was highly produced after 5 days of culture. Optimal pH and temperature conditions for the production of anti-VRSA substance were pH 7.0 and $28^{\circ}C$, respectively.

Changes in Skin Condition by Frequency of Sheet Mask Use (시트 마스크 사용에 따른 피부상태 변화 비교 연구)

  • Lee, Mi-Na
    • The Journal of the Korea Contents Association
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    • v.21 no.9
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    • pp.742-749
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    • 2021
  • This study aimed to comparatively analyze changes in skin conditions according to the frequency of sheet mask use among women in their 60s. An experiment was performed for 5 weeks from May 1 to June 3, 2020. A total of 70 women in their 60s, who understood the purposes of this study and agreed to the experiment were selected. All empirical analyses were conducted at a significance level of 5%, and the SPSS WIN 21 program was used to perform statistical analysis. Increased moisture content, decreased coarseness, changes in sebum levels, and decreased pigmentation, were significant effects of sheet mask use for 7days per week. As long as the sheet mask did not irritate the skin, had a positive effects on the skin the more often it was used. These results can help in enhancing the effectiveness of systematic measurement and improvement of skin type. To determine factors affecting skin conditions, it is necessary to investigate and compare factors such as effects of dietary or living habits, genetic influence, and living environments.