• Food Science and Biotechnology
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• 제15권3호
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• pp.447-452
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• 2006

To determine the functions of specific cell wall polysaccharides, polysaccharides of three mutants, mur3-1, mur3-2, and mur3-3, obtained from Arabidopsis wild type, underwent structural characterization. Upon sequential separation of pectins (RG-I and RG-II) and cross-linking glycans (xyloglucan, XG), only XG was affected by the mud mutation. Wild-type XG contained a considerable amount of fucose, whereas the fucose level in mur3 XGs was less than 20% that of wild type. Further analysis of XGs by matrix-assisted laser-induced/ionization time-of-flight (MALDI-TOF) mass spectrometry indicated that mud lines considerably or completely lost the fucosylated XG oligosaccharides such as XXFG and XLFG and the double-galactosylated oligosaccharide XLLG $^1H$-NMR spectroscopic analyses of the XG oligosaccharides from mur3-3 plant revealed the absence of fucose and a galactose level in the galactosylated side chain that was reduced by 40% compared to that of Arabidopsis wild-type plant. In contrast, 85% less fucose and a slight loss of galactose were observed in the mur3-1 and mur3-2 lines which show normal growth habit. Of the three Arabidopsis mur3 lines studied here, mur3-3 is disrupted by a T-DNA insertion in the exon of MUR3 which encodes XG-specific galactosyltransferase, and exhibits slight dwarfism. These results indicated that the T-DNA insertion at the MUR3 locus did not induce the complete loss of galactose in XG, and that galactose, rather than fucose, in the XG side chains made a major contribution to overall wall strength.

• 대한의용생체공학회:의공학회지
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• 제44권6호
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• pp.465-475
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• 2023

In this study, we demonstrated a silica nanofibrous membrane based on the electrospinning process and evaluated its DNA isolation and purification performance in PCR pretreatment. Generally, silica membranes made of non-woven fabric are used for PCR pretreatment, but this study aimed to improve the efficiency of the pretreatment process by developing a nanofiber-type silica membrane with high specific surface area and porosity. In order to manufacture a nanofiber-shaped silica film while maintaining the original physical properties of silica, nanofiber membranes produced by adding various concentrations of PEO (5 wt%, 8 wt%, and 10 wt%) to silica prepared by the sol-gel method were compared. In terms of nanofiber membrane production, the higher the PEO concentration, the more effective it was in producing nanofiber membranes. The produced silica nanofiber membrane was inserted to a pretreatment device used in commercial PCR equipment, and the pretreatment performance was compared and verified using Salmonella bacteria. When Salmonella was used, samples containing 5 wt% PEO showed superior PCR efficiency compared to samples containing 8 wt% and 10 wt% PEO. These results show that adding 5 wt% of PEO can effectively improve DNA purification and separation by producing a nanofiber-shaped silica film while maintaining the physical properties of silica. We expect that this study will contribute to the development of effective PCR pretreatment technology essential for various molecular biology applications.

• 생태와환경
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• 제56권1호
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• pp.94-103
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• 2023

휴면포자와 같은 남조류의 휴면세포는 남조류의 초기발생 및 대발생의 중요한 씨앗세포이다. 이러한 중요성으로 인해 퇴적층에 존재하는 휴면세포를 분리하기 위해서 다양한 방법들이 시도되었다. Ludox는 해양퇴적물의 세포분리에 주로 활용되는 용액이지만 담수에서는 정확한 사용법을 찾기 어렵다. 본 연구에서는 가장 많이 사용되는 두 가지 Ludox방법(퇴적물 직접처리, 퇴적물 증류수 현탁처리)을 비교하고, 담수 퇴적물에서 남조류 휴면세포의 분리 및 유전자 증폭 효율이 높은 방법을 제안하였다. 퇴적물에서 발견된 휴면세포는 대부분 염주말목의 휴면포자로써 Dolichospermum, Cylindrospermum, Aphanizomenon의 휴면포자 형태와 유사하였다. 퇴적물을 증류수에 현탁하여 처리한 시료보다 퇴적물 그대로 사용한 시료에서 20배 더 많은 휴면포자가 발견되었으며 증류수로 현탁된 퇴적물에서는 분리되지 않은 세포가 대부분 pellet 퇴적물 표층에서 발견되었다. Ludox를 통해 층 분리된 휴면포자는 수층의 특정 깊이에서 밀집하기보다는 주로 상층과 하층에 넓게 퍼져있었다. 퇴적물을 그대로 사용한 시료에서 mibC, Geo, 16S rDNA 유전자 모두 증폭산물이 확인되었으나 퇴적물을 증류수로 현탁한 시료에서는 모든 유전자의 증폭산물이 발견되지 않았다. 따라서 담수 퇴적물에서 남조류의 휴면세포를 분리하는 경우에는 5~10 g의 퇴적물을 전처리 없이 그대로 사용하며, 퇴적물량 4배 부피의 Ludox를 첨가할 때 세포 분리 및 유전자 증폭 효율이 높았다. 본 연구에서 제시하는 Ludox 처리방법은 담수퇴적층에 존재하는 남조류 휴면세포를 분리하기 위한 방법으로써 다른 생물군에서는 동일한 효율이 나타나지 않을 수 있다. 따라서 다른 생물군의 분리에 Ludox를 적용하기 위해서는 퇴적물 전처리 방법 및 대상생물이 존재하는 수층을 파악하는 사전실험이 반드시 필요하다.

• Journal of Microbiology
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• 제44권6호
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• pp.600-606
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• 2006

Spore forming actinobacteria (sporoactinobacteria) isolated from soils with an acidic pH in Pinus thunbergii forests and coal mine waste were subjected to taxonomic characterization. For the isolation of acidophilic actinobacteria, acidified starch casein agar (pH adjusted to 4-5) was used. The numbers of actinobacteria growing in acidic media were between $3.2{\times}10^4$ and $8.0{\times}10^6$ CFU/g soil. Forty three acidophilic actinobacterial strains were isolated and their 16S rDNA sequences were determined. The isolates were divided into eight distinctive phylogenetic clusters within the variation encompassed by the family Streptomycetaceae. Four clusters among them were assigned to the genus Streptacidiphilus, whereas the remaining four were assigned to Streptomyces. The clusters belonging to either Streptomyces or Streptacidiphilus did not form a monophyletic clade. The growth pH profiles indicated that the representative isolates grew best between pH 5 and 6. It is evident from this study that acidity has played a critical role in the differentiation of the family Streptomycetaceae, and also that different mechanisms might have resulted in the evolution of two groups, Streptacidiphilus (strict acidophiles) and neutrotolerant acidophilic Streptomyces. The effect of geographic separation was clearly seen among the Streptacidiphilus isolates, which may be a key factor in speciation of the genus.

• BMB Reports
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• 제28권4호
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• pp.359-362
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• 1995

The highly polymorphic variable number of tandem repeat (VNTR) loci in the human genome are informative markers for the genetic characterization of individuals in the paternity test and forensic science as well as for the study of human disease. In this study, VNTR loci D1S80 and D2S123 have been amplified by PCR and the amplified length polymorphic alleles were detected with a discontinuous vertical PAGE system and silver staining. For explicit DNA typing, PCR optimization, in which amplification efficiencies are similar over a wide range of allele sizes, non-specific amplifications are minimal, and new longer alleles have high amplification efficiency, has been performed by changing the PCR reaction buffer composition and thermal cycling conditions. It turned out that adding an appropriate amount of Tween 20 and NP40 to the PCR reaction buffer and raising the annealing temperature to $68^{\circ}C$ in thermal cycling made it possible for optimal VNTR loci amplification. A modified PAGE system for VNTR separation was established. Under these conditions, new longer alleles in the 01580 locus were discovered and 025123 pattern changes in colorectal tumors were observed. These technical tips are valuable for detecting various amplified fragment length polymorphisms.

• Bulletin of the Korean Chemical Society
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• 제29권1호
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• pp.153-158
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• 2008

A microchip-based capillary gel electrophoresis (MCGE) technique was developed for the ultra-fast detection and differentiation of Candidatus Mycoplasma haemominutum (Candidatus M. haemominutum, California strain) and Mycoplasma haemofelis (M. haemofelis, Ohio strain) in Korean feral cats through the application of programmed field strength gradients (PFSG) in a conventional glass double-T microchip. The effects of the poly (ethyleneoxide) (PEO) concentration and electric field strength on the separation of DNA fragments were investigated. The PCR-amplified products of Candidatus M. haemominutum (202-bp) and M. haemofelis (273-bp) were analyzed by MCGE within 75 s under a constant applied electric field of 117.6 V/cm and a sieving matrix of 0.3% PEO (Mr 8 000 000). When the PFSG was applied, MCGE analysis generated results 6.8-times faster without any loss of resolution or reproducibility. The MCGE-PFSG technique was also applied to eleven samples selected randomly from 33 positive samples. The samples were detected and differentiated within 11 s. The analysis time of the MCGE-PFSG technique was approximately 980-times faster than that using conventional slab gel electrophoresis.

• ALGAE
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• 제30권2호
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• pp.89-101
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• 2015

Ecklonia cava Kjellman is a common kelp found in shallow subtidal in warm-temperate waters in the northwest Pacific Ocean. This species has shown substantial morphological variation along with subsistence in different locations and local environments. We quantified the magnitude of morphological variation of E. cava from six populations along ~700 km of coastline from Jeju Island to Dokdo in Korea. In addition, we examined genetic distance among the populations using random amplified polymorphic DNA (RAPD) analysis. Most morphological characteristics investigated were significantly different among locations. Multivariate analyses indicated two phenetically distinct groups (nearshore, sheltered vs. offshore, exposed), indicating wave exposure with turbidity are presumably major factors for the separation. With RAPD data, results of Nei's diversity (H) and AMOVA showed considerable variations in within- and between-populations. Pairwise ${\Phi}_{ST}$ and $N_m$ values indicated moderate gene flow between the six locations. Results of Nei's analysis revealed three genetically distinct groups, not consistent with the morphological groupings, indicating that a time gap may exist between morphological and genetic variations. This study also suggests dispersal distance of this kelp may be longer than what is commonly thought and genetic similarity in the populations was largely reflected by the direction of ocean current rather than just geographical distance.

• Applied Biological Chemistry
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• 제40권2호
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• pp.101-106
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• 1997

발효시켜서 만드는 감식초에서 시료를 채취하여 배양하고, pellicle을 형성하는 single colony를 cellulose 생합성균으로 분리하였다. 분리된 균주는 형태적 특성, 알콜의 재산화 등의 생리, 생화학적 특성 등에 의하여 Acetobacter속으로 분류되었으며, Acetobacter CBI-2라고 명명하였다. Acetobacter CBI-2는 정치배양 시에 기존의 생합성 균주로 알려진 Acetobacter xylinum과 대등한 cellulose 생합성 능력을 나타내었다. Acetobacter CBI-2이 생성한 고분자물질의 분해산물을 TLC법으로 확인한 결과 기존 cellulose의 것과 일치하였다. Acetobacter CBI-2에서 genomic DNA를 분리 정제하여 cel A 영역을 probe로하여 hybridization 시킨 결과, 상동성을 나타내어 분리 균주에는 cellulose 생합성 관련 유전자가 존재함을 나타내었다.

• 대한화학회지
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• 제37권3호
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• pp.355-363
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• 1993

Whatmann chromatography paper(3MM)을 지지체로 사용하여 phosphite-triester법에 의해 미량의 염기가 부착된 5'-d(GAATTCCGGCCA)와 5'-d(GGAGCTGTC)의 두 oligodeoxyribonucleotide를 합성 하였다. Trityl yield 분석에 의하면 76.1${\%}$와 86.5${\%}$의 평균 coupling 수율을 각각 얻었다.두 oligodeoxyribonucleotide들을 HPLC를 이용하여 분리 정제할 수 있었고 snake venom phosphodiesterase와 bacterial alkaline phosphatase에 의해 분리 정량화할 수 있었으며 이때의 염기의 수는 합성을 시도한 염기수와 일치했다. 이처럼 paper를 지지체로 이용한 oligodeoxyribonucleotide의 합성은 값싸고 동시에 많은 염기가 부착된 미량의 DNA를 빠르고 짧은 기간에 수행할 수 있는 장점이 있다.

• Journal of the Korean Wood Science and Technology
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• 제50권1호
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• pp.12-30
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• 2022

Many studies on plant extracts have been reported for the treatment of candidiasis caused by Candida albicans, a representative fungal infection. This study demonstrates the synergistic antifungal activity of the combination of Phellodendri Cortex and Magnoliae Cortex, previously reported to have antifungal efficacy. Considering the antifungal efficacy and the separation of the active constituents, berberine and magnolol, hot water extraction and carbon dioxide supercritical extraction were selected for Phellodendri Cortex and Magnoliae Cortex, respectively. A combination of 0.55 g/L hot water extract of Phellodendri Cortex and 0.59 g/L carbon dioxide supercritical extract of Magnoliae Cortex showed synergistic antifungal activity. The synergistic antifungal activity of 160 μM berberine and 100 μM magnolol, which are representative antifungal compounds of Phellodendri Cortex and Magnoliae Cortex, respectively, contributes to the synergistic antifungal effect of their extracts. The additive decrease in cellular ergosterol level and the increased antifungal efficacy by extracellular ergosterol suggest that disruption of the biological function of ergosterol in the cell membrane is not responsible for the synergistic antifungal activity of berberine and magnolol. Synergistic cellular release of chromosomal DNA upon mixing berberine and magnolol indicates that disruption of the cellular structure is responsible for the synergistic antifungal effect of berberine and magnolol.