• Title/Summary/Keyword: molecular biological techniques

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Search Space Reduction Techniques in Small Molecular Docking (소분자 도킹에서 탐색공간의 축소 방법)

  • Cho, Seung Joo
    • Journal of Integrative Natural Science
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    • v.3 no.3
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    • pp.143-147
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    • 2010
  • Since it is of great importance to know how a ligand binds to a receptor, there have been a lot of efforts to improve the quality of prediction of docking poses. Earlier efforts were focused on improving search algorithm and scoring function in a docking program resulting in a partial improvement with a lot of variations. Although these are basically very important and essential, more tangible improvements came from the reduction of search space. In a normal docking study, the approximate active site is assumed to be known. After defining active site, scoring functions and search algorithms are used to locate the expected binding pose within this search space. A good search algorithm will sample wisely toward the correct binding pose. By careful study of receptor structure, it was possible to prioritize sub-space in the active site using "receptor-based pharmacophores" or "hot spots". In a sense, these techniques reduce the search space from the beginning. Further improvements were made when the bound ligand structure is available, i.e., the searching could be directed by molecular similarity using ligand information. This could be very helpful to increase the accuracy of binding pose. In addition, if the biological activity data is available, docking program could be improved to the level of being useful in affinity prediction for a series of congeneric ligands. Since the number of co-crystal structures is increasing in protein databank, "Ligand-Guided Docking" to reduce the search space would be more important to improve the accuracy of docking pose prediction and the efficiency of virtual screening. Further improvements in this area would be useful to produce more reliable docking programs.

Developing a Bioinformatics Tool for Peptide Nucleic Acid (PNA) antisense Technique Utilizing Parallel Computing System (Peptide Nucleic Acid(PNA)를 이용한 antisense 기법에 적용할 병렬 컴퓨팅용 Bioinformatics tool 개발)

  • Kim Seong-Jo;Jeon Ho-Sang;Hong Seung-Pyo;Kim Hyon-Chang;Kim Han-Jip;Min Churl-K
    • Proceedings of the Korean Information Science Society Conference
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    • 2006.06a
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    • pp.43-45
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    • 2006
  • Unlike RNA interference, whose usage is limited to eukaryotic cells, Peptide Nucleic Acid (PNA) technique is applicable to both eukaryotic and prokaryotic cells. PNA has been proven to be an effective agent for blocking gene expressions and has several advantages over other antisense techniques. Here we developed a parallel computing software that provides the ideal sequences to design PNA oligos to prevent any off-target effects. We applied a new approach in our location-finding algorithm that finds a target gene from the whole genome sequence. Message Passing Interface (MPI) was used to perform parallel computing in order to reduce the calculation time. The software will help biologists design more accurate and effective antisense PNA by minimizing the chance of off-target effects.

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Molecular and Cellular Studies of Seed Storage Proteins from Rice and Wheat

  • Kim, Woo-Taek
    • Applied Biological Chemistry
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    • v.32 no.1
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    • pp.64-72
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    • 1989
  • Near full length cDNA clones encoding the rice seed storage protein, prolamine, were isolated and divided into two homology classes based on cross-hybridization and DNA sequencing analysis. These cDNA clones contain a single open reading frame encoding a putative rice prolamine precursor(M.W.=17,200) possessing atypical 14 amino acid signal peptide. Clones of these two homology classes diverge mainly by insertions/deletions of short nucleotide stretches and point mutations. The deduced primary structures of both types of prolamine polypeptides are devoid of any major tandem repetitive sequences, a feature prevalent in other cereal prolamines. No significant homology teas detected between the rice prolamine and other cereal prolamines, indicating that the rice gene evolved from a different ancestor that gave rise to other cereal prolamine genes. Developing wheat and rice endosperms were examined using ultrathin sections prepared from tissues harvested at various days after flowering. By immunocytochemical localization techniques, wheat prolamines are localized within vesicles from Golgi apparatus and in homogeneous regions of protein bodies. The involvement of the goli apparatus in the packaging of wheat prolamines into protein bodies indicates a pathway which differs from the mode of other cereal prolamines and resembles the mechanism employed for the storage of rice glutelin and legume globulins.

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An Investigation of the Terahertz Absorption Characteristics of a Graphene Oxide Aqueous Solution Using Microfluidic Technology

  • Ningyi Cai;Boyan Zhang;Qinghao Meng;Siyu Qian;Bo Su;Hailin Cui;Shengbo Zhang;Cunlin Zhang
    • Current Optics and Photonics
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    • v.7 no.2
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    • pp.119-126
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    • 2023
  • The vibratory and rotational levels of many biological macromolecules lie in the terahertz (THz) band, which means that THz techniques can be used to identify and detect them. Moreover, since the biological activity of most biomolecules only becomes apparent in aqueous solution, we use microfluidic technology to study the biological properties of these biomolecules. THz time-domain spectroscopy was used to study the THz absorption characteristics of graphene oxide (GO) aqueous solution at different concentrations and different exposure times in fixed electric or magnetic fields. The results show that the spectral characteristics of the GO solution varied with the concentration: as the concentration increased, the THz absorption decreased. The results also show that after placing the solution in an external electric field, the absorption of THz first increased and then decreased. When the solution was placed in a magnetic field, the THz absorption increased with the increase in standing time. In this paper, these results are explained based on considerations of what is occurring at the molecular scale. The results of this study provide technical support for the further study of GO and will assist with its improved application in various fields.

Fabrication of Tissue Engineered Intervertebral Disc Using Enable 3D bio-printing and Scaffod-Free technologies (3D 바이오프린팅과 무지지체 조직공학 기술 기반 추간판 복합 조직 제작)

  • Kim, Byeong Kook;Park, Jinho;Park, Sang-Hyug
    • Journal of Biomedical Engineering Research
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    • v.39 no.1
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    • pp.22-29
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    • 2018
  • Intervertebral disc(IVD) mainly consists of Annulus fibrosus(AF) and Nucleus pulposus(NP), playing a role of distributing a mechanical load on vertebral body. IVD tissue engineering has been developed the methods to achieve anatomic morphology and restoration of biological function. The goal of present study is to identify the possibilities for creating a substitute of IVD the morphology and biological functions are the same as undamaged complete IVD. To fabricate the AF and NP combine biphasic IVD tissue, AF tissue scaffolds have been printed by 3D bio-printing system with natural biomaterials and NP tissues have been prepared by scaffold-free culture system. We evaluated whether the combined structure of 3D printed AF scaffold and scaffold-free NP tissue construct could support the architecture and cell functions as IVD tissue. 3D printed AF scaffolds were printed with 60 degree angle stripe patterned lamella structure(the inner-diameter is 5mm, outer-diameter is 10 mm and height is 3 mm). In the cytotoxicity test, the 3D printed AF scaffold showed good cell compatibility. The results of histological and immunohistochemical staining also showed the newly synthesized collagens and glycosaminoglycans, which are specific makers of AF tissue. And scaffold-free NP tissue actively synthesized glycosaminoglycans and type 2 collagen, which are the major components of NP tissue. When we combined two engineered tissues to realize the IVD, combined biphasic tissues showed a good integration between the two tissues. In conclusion, this study describes the fabrication of Engineered biphasic IVD tissue by using enable techniques of tissue engineering. This fabricated biphasic tissue would be used as a model system for the study of the native IVD tissue. In the future, it may have the potential to replace the damaged IVD in the future.

Status of Molecular Biotechnology Research Based on Tissue Culture of Soybean (콩 조직배양 기술에 기반한 생명공학 연구 동향)

  • Seo, Mi-Suk;Cho, Chuloh;Choi, Man-Soo;Chun, JaeBuhm;Jin, Mina;Kim, Dool-Yi
    • Korean Journal of Plant Resources
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    • v.33 no.5
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    • pp.536-549
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    • 2020
  • Soybean (Glycine max (L.) Merrill) is one of the most important crops of the world. With the completion of the soybean genome sequence, the Korean soybean core collection consisted of 430 accessions with genetic and phenotypic diversity was constructed in recent year. The availability of genome sequences and core collection will result in the crop improvement by molecular breeding using the various accessions and genome editing approaches. Efficient tissue culture techniques, such as haploid production, protoplast culture and plant regeneration from various organs are essential for the successful molecular biological approach and crop improvement. However, soybean is still considered to be recalcitrant in tissue culture because of the low frequency of regeneration and limitation of available responsive cultivars. In this study, we discuss the recent studies of tissue culture technology and methodology for efficient tissue culture to genetic improvement and application of molecular biotechnology in soybean.

Molecular Identification of Adoxophyes honmai (Yasuda) (Lepidoptera: Tortricidae) Based on Mitochondrial COI Gene Sequences

  • Lee, So Young;Park, Hyungjin;Boo, Kyung Saeng;Park, Kyu-Tek;Cho, Soowon
    • Molecules and Cells
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    • v.19 no.3
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    • pp.391-397
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    • 2005
  • Molecular identification techniques are used where morphological characters are not useful for distinguishing species that resemble each other closely. The example studied here is the Adoxophyes species complex, in which A. orana (Fischer von $R{\ddot{o}}sslerstamm$) is officially the only known Korean species in the genus Adoxophyes (Lepidoptera: Tortricidae). However there have been suspicions that at least two types of A. orana exist in Korea based on the distribution and range of the host, with A. orana attacking apples and peaches, and another Adoxophyes sp. attacking tea and pears. The latter is presumed to be A. honmai (Yasuda), but the two have remained confused because of their extreme morphological similarity, despite several Asian studies of pheromonal and morphological characteristics. To confirm the occurrence of an Adoxophyes species other than A. orana in Korea, we compared 940 bp of the mitochondrial cytochrome oxidase I (COI) gene from 16 samples of Adoxophyes and found that there is a second Adoxophyes species different from A. orana. Comparison of the different sequences to that of Japanese A. honmai confirmed that they belong to the latter. From the sequence difference between the two Korean species, we were able to develop new PCR primer sets that distinguish them. This molecular identification technique with no enzyme digestion or sequencing step is a convenient and rapid way of differentiating between species that are hard to distinguish morphologically.

Application of Molecular Biology to Rumen Microbes -Review-

  • Kobayashi, Y.;Onodera, R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.1
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    • pp.77-83
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    • 1999
  • Molecular biological techniques that recently developed, have made it possible to realize some of new attempts in the research field of rumen microbiology. Those are 1) cloning of genes from rumen microorganisms mainly in E. coli, 2) transformation of rumen bacteria and 3) ecological analysis with nonculturing methods. Most of the cloned genes are for polysaccharidase enzymes such as endoglucanase, xylanase, amylase, chitinase and others, and the cloning rendered gene structural analyses by sequencing and also characterization of the translated products through easier purification. Electrotransformation of Butyrivibrio fibrisolvens and Prevotella ruminicola have been made toward the direction for obtaining more fibrolytic, acid-tolerant, depoisoning or essential amino acids-producing rumen bacterium. These primarily required stable and efficient gene transfer systems. Some vectors, constructed from native plasmids of rumen bacteria, are now available for successful gene introduction and expression in those rumen bacterial species. Probing and PCR-based methodologies have also been developed for detecting specific bacterial species and even strains. These are much due to accumulation of rRNA gene sequences of rumen microbes in databases. Although optimized analytical conditions are essential to reliable and reproducible estimation of the targeted microbes, the methods permit long term storage of frozen samples, providing us ease in analytical work as compared with a traditional method based on culturing. Moreover, the methods seem to be promissing for obtaining taxonomic and evolutionary information on all the rumen microbes, whether they are culturable or not.

Meroparamycin Production by Newly Isolated Streptomyces sp. Strain MAR01: Taxonomy, Fermentation, Purification and Structural Elucidation

  • El-Naggar Moustafa Y.;El-Assar Samy A.;Abdul-Gawad Sahar M.
    • Journal of Microbiology
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    • v.44 no.4
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    • pp.432-438
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    • 2006
  • Twelve actinomycete strains were isolated from Egyptian soil. The isolated actinomycete strains were then screened with regard to their potential to generate antibiotics. The most potent of the producer strains was selected and identified. The cultural and physiological characteristics of the strain identified. the strain as a member of the genus Streptomyces. The nucleotide sequence of the 16S rRNA gene (1.5kb) of the most potent strain evidenced a 99% similarity with Streptomyces spp. and S. aureofaciens 16S rRNA genes, and the isolated strain was ultimately identified as Streptomyces sp. MAR01. The extraction of the fermentation broth of this strain resulted in the isolation of one major compound, which was active in vitro against gram-positive, gram-negative representatives and Candida albicans. The chemical structure of this bioactive compound was elucidated based on the spectroscopic data obtained from the application of MS, IR, UV, $^1H$ NMR, $^{13}C$ NMR, and elemental analysis techniques. Via comparison to the reference data in the relevant literature and in the database search, this antibiotic, which had a molecular formula of $C_{19}H_{29}NO_2$ and a molecular weight of 303.44, was determined to differ from those produced by this genus as well as the available known antibiotics. Therefore, this antibiotic was designated Meroparamycin.

Detection and Analysis of DNA Hybridization Characteristics by using Thermodynamic Method (열역학법을 이용한 DNA hybridization 특성 검출 및 해석)

  • Kim, Do-Gyun;Gwon, Yeong-Su
    • The Transactions of the Korean Institute of Electrical Engineers C
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    • v.51 no.6
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    • pp.265-270
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    • 2002
  • The determination of DNA hybridization reaction can apply the molecular biology research, clinic diagnostics, bioengineering, environment monitoring, food science and application area. So, the improvement of DNA hybridization detection method is very important for the determination of this hybridization reaction. Several molecular biological techniques require accurate predictions of matched versus mismatched hybridization thermodynamics, such as PCR, sequencing by hybridization, gene diagnostics and antisense oligonucleotide probes. In addition, recent developments of oligonucleotide chip arrays as means for biochemical assays and DNA sequencing requires accurate knowledge of hybridization thermodynamics and population ratios at matched and mismatched target sites. In this study, we report the characteristics of the probe and matched, mismatched target oligonucleotide hybridization reaction using thermodynamic method. Thermodynamic of 5 oligonucleotides with central and terminal mismatch sequences were obtained by measured UV-absorbance as a function of temperature. The data show that the nearest-neighbor base-pair model is adequate for predicting thermodynamics of oligonucleotides with average deviations for $\Delta$H$^{0}$ , $\Delta$S$^{0}$ , $\Delta$G$_{37}$ $^{0}$ and T$_{m}$, respectively.>$^{0}$ and T$_{m}$, respectively.