• 제목/요약/키워드: Systems Biotechnology

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Applications of DNA Microarray in Disease Diagnostics

  • Yoo, Seung-Min;Choi, Jong-Hyun;Lee, Sang-Yup;Yoo, Nae-Choon
    • Journal of Microbiology and Biotechnology
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    • 제19권7호
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    • pp.635-646
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    • 2009
  • Rapid and accurate diagnosis of diseases is very important for appropriate treatment of patients. Recent advances in molecular-level interaction and detection technologies are upgrading the clinical diagnostics by providing new ways of diagnosis, with higher speed and accuracy. In particular, DNA microarrays can be efficiently used in clinical diagnostics which span from discovery of diseaserelevant genes to diagnosis using its biomarkers. Diagnostic DNA microarrays have been used for genotyping and determination of disease-relevant genes or agents causing diseases, mutation analysis, screening of single nucleotide polymorphisms (SNPs), detection of chromosome abnormalities, and global determination of posttranslational modification. The performance of DNA-microarray-based diagnosis is continuously improving by the integration of other tools. Thus, DNA microarrays will play a central role in clinical diagnostics and will become a gold standard method for disease diagnosis. In this paper, various applications of DNA microarrays in disease diagnosis are reviewed. Special effort was made to cover the information disclosed in the patents so that recent trends and missing applications can be revealed.

Single-Base Genome Editing in Corynebacterium glutamicum with the Help of Negative Selection by Target-Mismatched CRISPR/Cpf1

  • Kim, Hyun Ju;Oh, Se Young;Lee, Sang Jun
    • Journal of Microbiology and Biotechnology
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    • 제30권10호
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    • pp.1583-1591
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    • 2020
  • CRISPR/Cpf1 has emerged as a new CRISPR-based genome editing tool because, in comparison with CRIPSR/Cas9, it has a different T-rich PAM sequence to expand the target DNA sequence. Single-base editing in the microbial genome can be facilitated by oligonucleotide-directed mutagenesis (ODM) followed by negative selection with the CRISPR/Cpf1 system. However, single point mutations aided by Cpf1 negative selection have been rarely reported in Corynebacterium glutamicum. This study aimed to introduce an amber stop codon in crtEb encoding lycopene hydratase, through ODM and Cpf1-mediated negative selection; deficiency of this enzyme causes pink coloration due to lycopene accumulation in C. glutamicum. Consequently, on using double-, triple-, and quadruple-base-mutagenic oligonucleotides, 91.5-95.3% pink cells were obtained among the total live C. glutamicum cells. However, among the negatively selected live cells, 0.6% pink cells were obtained using single-base-mutagenic oligonucleotides, indicating that very few single-base mutations were introduced, possibly owing to mismatch tolerance. This led to the consideration of various target-mismatched crRNAs to prevent the death of single-base-edited cells. Consequently, we obtained 99.7% pink colonies after CRISPR/Cpf1-mediated negative selection using an appropriate single-mismatched crRNA. Furthermore, Sanger sequencing revealed that single-base mutations were successfully edited in the 99.7% of pink cells, while only two of nine among 0.6% of pink cells were correctly edited. The results indicate that the target-mismatched Cpf1 negative selection can assist in efficient and accurate single-base genome editing methods in C. glutamicum.

Factors for high frequency plant regeneration in tissue cultures of Indian mustard (Brassica juncea L.)

  • Bhuiyan, Mohammed Shafi Ullah;Min, Sung-Ran;Choi, Kwan-Sam;Lim, Yong-Pyo;Liu, Jang-Ryol
    • Journal of Plant Biotechnology
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    • 제36권2호
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    • pp.137-143
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    • 2009
  • An efficient system for high frequency plant regeneration was established through investigating various factors such as plant growth regulator combinations, explant types and ages, and addition of $AgNO_3$ influenced on shoot regeneration in Brassica juncea L. cv. BARI sarisha-10. Murashige and Skoog (MS) medium supplemented with 0.1 mg/L NAA (1-naphthaleneacetic acid) and 1 mg/L BA (6-benzyladenine) showed the maximum shoot regeneration frequency (56.67%) among the different combinations of NAA and BA. Explant type, explant age, and addition of $AgNO_3$ also significantly affected shoot regeneration. Of the four type of explants (cotyledon, hypocotyl, root, and leaf explants)- cotyledon explants produced the highest shoot regeneration frequency and hypocotyls explants produced the highest number of shoots per explant, whereas root explants did not produce any shoot. The cotyledonary explants from Four-day-old seedlings showed the maximum shoot regeneration frequency and number of shoots per explant. Shoot regeneration frequency increased significantly by adding $AgNO_3$ to the medium. Two mg/L $AgNO_3$ appeared to be the best for shoot regeneration with the highest shoot regeneration frequency (86.67%) and number of shoots per explant (7.5 shoots). Considerable variation in shoot regeneration from cotyledonay explants was observed within the B. juncea L. genotypes. The shoot regeneration frequency ranged from 47.78% for cv. Shambol to 91.11% for cv. Rai-5. In terms of the number of shoots produced per explant, B. juncea L. cv. Daulot showed the maximum efficiency. MS medium supplemented with 0.1 mg/L NAA showed the highest frequency of rooting. The regenerated plantlets were transferred to pot soil and grown to maturity in the greenhouse. All plants were fertile and morphologically identical with the source plants.

Report on 30 unrecorded bacterial species of the phylum Firmicutes isolated from Korea in 2016

  • Nahar, Shamsun;Lee, Do-Hoon;Bae, Jin-Woo;Im, Wan-Taek;Jahng, Kwang Yeop;Joh, Kiseong;Kim, Wonyong;Lee, Soon Dong;Yi, Hana;Cha, Chang-Jun
    • Journal of Species Research
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    • 제7권1호
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    • pp.50-59
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    • 2018
  • During the course of investigation of indigenous prokaryotic species in Korea, a total of 30 bacterial strains belonging to the phylum Firmicutes were isolated from diverse environmental sites such as soil, avian feces, wastewater treatment plants, fermented vegetables, seawater, algae, sea cucumber, octopus and tidal flat sediment. Phylogenetic analysis based on 16S rRNA gene sequences revealed that each strain showed high sequence similarity (${\geq}98.7%$) to the closest type strain and formed a robust phylogenetic clade with the most closely related species in the phylum Firmicutes. To date, there is no official record of these 30 species in Korea. Therefore, we report 26 species of 12 genera in the order Bacillales and 4 species of 4 genera in the order Lactobacillales which have not been reported in Korea. Morphological and biochemical characteristics, isolation sources and NIBR deposit numbers are described in the species descriptions.

FT-IR 스펙트럼 데이터의 다변량 통계분석 기법을 이용한 바위솔속 식물의 분류학적 유연관계 예측 및 판별 (Prediction and discrimination of taxonomic relationship within Orostachys species using FT-IR spectroscopy combined by multivariate analysis)

  • 권용국;김석원;서정민;우태하;유장렬
    • Journal of Plant Biotechnology
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    • 제38권1호
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    • pp.9-14
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    • 2011
  • To determine whether pattern recognition based on metabolite fingerprinting for whole cell extracts can be used to discriminate cultivars metabolically, leaves of nine commercial Orostachys plants were subjected to Fourier transform infrared spectroscopy (FT-IR). FT-IR spectral data from leaves were analyzed by principal component analysis (PCA) and Partial least square discriminant analysis (PLS-DA). The dendrogram based on hierarchical clustering analysis of these PLS-DA data separated the nine Orostachys species into five major groups. The first group consisted of O. iwarenge 'Yimge', 'Jeju', 'Jeongsun' and O. margaritifolius 'Jinju' whereas in the second group, 'Sacheon' was clustered with 'Busan,' both of which belong to O. malacophylla species. However, 'Samchuk', belong to O. malacophylla was not clustered with the other O. malacophylla species. In addition, O. minuta and O. japonica were separated to the other Orostachys plants. Thus we suggested that the hierarchical dendrogram based on PLS-DA of FT-IR spectral data from leaves represented the most probable chemotaxonomical relationship between commercial Orostachys plants. Furthermore these metabolic discrimination systems could be applied for reestablishment of precise taxonomic classification of commercial Orostachys plants.

Metagenomic SMRT Sequencing-Based Exploration of Novel Lignocellulose-Degrading Capability in Wood Detritus from Torreya nucifera in Bija Forest on Jeju Island

  • Oh, Han Na;Lee, Tae Kwon;Park, Jae Wan;No, Jee Hyun;Kim, Dockyu;Sul, Woo Jun
    • Journal of Microbiology and Biotechnology
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    • 제27권9호
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    • pp.1670-1680
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    • 2017
  • Lignocellulose, composed mostly of cellulose, hemicellulose, and lignin generated through secondary growth of woody plant, is considered as promising resources for biofuel. In order to use lignocellulose as a biofuel, biodegradation besides high-cost chemical treatments were applied, but knowledge on the decomposition of lignocellulose occurring in a natural environment is insufficient. We analyzed the 16S rRNA gene and metagenome to understand how the lignocellulose is decomposed naturally in decayed Torreya nucifera (L) of Bija forest (Bijarim) in Gotjawal, an ecologically distinct environment. A total of 464,360 reads were obtained from 16S rRNA gene sequencing, representing diverse phyla; Proteobacteria (51%), Bacteroidetes (11%) and Actinobacteria (10%). The metagenome analysis using single molecules real-time sequencing revealed that the assembled contigs determined originated from Proteobacteria (58%) and Actinobacteria (10.3%). Carbohydrate Active enZYmes (CAZy)- and Protein families (Pfam)-based analysis showed that Proteobacteria was involved in degrading whole lignocellulose, and Actinobacteria played a role only in a part of hemicellulose degradation. Combining these results, it suggested that Proteobacteria and Actinobacteria had selective biodegradation potential for different lignocellulose substrates. Thus, it is considered that understanding of the systemic microbial degradation pathways may be a useful strategy for recycle of lignocellulosic biomass, and the microbial enzymes in Bija forest can be useful natural resources in industrial processes.

Low-Temperature Sintering Behavior of Aluminum Nitride Ceramics with Added Copper Oxide or Copper

  • Hwang, Jin-Geun;Oh, Kyung-Sik;Chung, Tai-Joo;Kim, Tae-Heui;Paek, Yeong-Kyeun
    • 한국세라믹학회지
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    • 제56권1호
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    • pp.104-110
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    • 2019
  • The low-temperature sintering behavior of AlN was investigated through a conventional method. $CaF_2$, CuO and Cu were selected as additives based on their low melting points. When sintered at $1600^{\circ}C$ for 8 h in $N_2$ atmosphere, a sample density > 98% was obtained. The X-ray data indicated that eutectic reactions below $1200^{\circ}C$ were found. Therefore, the current systems have lower liquid formation temperatures than other systems. The liquid phase showed high dihedral angles at triple grain junctions, indicating that the liquid had poor wettability on the grain surfaces. Eventually, the liquid was likely to vaporize due to the unfavorable wetting condition. As a result, a microstructure with clean grain boundaries was obtained, resulting in higher contiguity between grains. From EDS analysis, oxygen impurity seems to be well removed in AlN lattice. Therefore, it is believed that the current systems are beneficial for reducing sintering temperature and improving oxygen removal.

Mycosporine-like Amino Acids as Natural Scavengers of Singlet Oxygen in Marine Organisms: Photoprotection of Biological Systems

  • Suh, Hwa-Jin;Lee, Hyun--Woo;Jin Jung
    • Journal of Photoscience
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    • 제9권2호
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    • pp.63-65
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    • 2002
  • This report concerns a putative role of mycosporine-like amino acids (MAA) as natural scavengers of singlet oxygen ($^1$O$_2$) in marine organisms. MAA prepared from the ascidian Lissoclinum patella were found to protect biological systems against detrimental effects of the type II photosensitization in vitro. L. patella MAA were resolved into five components, and the relative $^1$0$_2$ quenching efficiencies were measured for three major components in aqueous media. It turned out that they were all effective in scavenging $^1$0$_2$, to different degrees albeit. The results suggest that physiological relevance of MAA in marine organisms may be found in a 'built-in' defense against photooxidative effects of sunlight.

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Current status on applications of conventional breeding techniques and biotechnological system in ornamentals

  • Kim, Jong Bo
    • Journal of Plant Biotechnology
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    • 제47권2호
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    • pp.107-117
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    • 2020
  • Flower industry is now growing due to the development of economy in many countries. Simultaneously, needs from consumers in flower market are varied widely. To satisfy the needs from consumers and deal with a variety of diseases from a lots of pathogens as well as climate change, new elite flower cultivars should be released in flower market. For this purpose, conventional and biotechnological techniques can be employed to make good cultivar. Therefore, this review describes the general overview of flower breeding techniques including cross-hybridization, mutation breeding and genetic transformation systems. Also, breeding systems for ornamentals derived from plant tissue culture such as embryo culture, in vitro fertilization, ovary/ovule culture and haploid production were reviewed. Furthermore, in this study recent development of the generation of new flower cultivars using marker-assisted breeding, plant transformation including particle bombardment and Agrobacterium tumefaciens as well as genome-editing technology were described. This review will be contributed to the development and releasement of new flower cultivars with horticulturally useful traits in the future.

Design of Bacterial Vector Systems for the Production of Recombinant Proteins in Escherichia coli

  • Mergulhao;Filipe J.M.;Gabriel A. Monteiro;Joaquim M.S. Cabral;M. Angela Taipa
    • Journal of Microbiology and Biotechnology
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    • 제14권1호
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    • pp.1-14
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    • 2004
  • More than twenty years have passed since the approval of the first recombinant DNA product for therapeutic use (recombinant human insulin, 1982). However, the biotechnology industry is still facing a shortage of manufacturing capacity due to the increasing demand of therapeutic proteins. This demand has prompted the search for a growing number of biological production systems but, nevertheless, the Gram-negative bacterium Escherichia coli remains one of the most attractive production hosts. This review highlights the most important features and developments of plasmid vector design, emphasizing the different reported strategies for improving the expression and secretion of heterologous proteins using the cellular machinery of E. coli.