• Title/Summary/Keyword: Cell manipulation

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Factors Influencing the Efficiency of In Vitro Embryo Production in the Pig

  • Lin, Tao;Lee, Jae Eun;Shin, Hyun Young;Oqani, Reza K.;Jin, Dong Il
    • Reproductive and Developmental Biology
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    • v.39 no.2
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    • pp.29-36
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    • 2015
  • Pigs are considered an ideal source of human disease model due to their physiological similarities to humans. However, the low efficiency of in vitro embryo production (IVP) is still a major barrier in the production of pig offspring with gene manipulation. Despite ongoing advances in the associated technologies, the developmental capacity of IVP pig embryos is still lower than that of their in vivo counterparts, as well as IVP embryos of other species (e.g., cattle and mice). The efficiency of IVP can be influenced by many factors that affect various critical steps in the process. The previous relevant reviews have focused on the in vitro maturation system, in vitro culture conditions, in vitro fertilization medium, issues with polyspermy, the utilized technologies, etc. In this review, we concentrate on factors that have not been fully detailed in prior reviews, such as the oocyte morphology, oocyte recovery methods, denuding procedures, first polar body morphology and embryo quality.

Construction and Transformation of an Endogenous Plasmid pBL1-free Brevibacterium lactofermentum (내재형 Plasmid pBL1이 제거된 Brevibacterium lactofermentum 개발과 형질전환)

  • 이규남;민본홍;윤기홍
    • Microbiology and Biotechnology Letters
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    • v.23 no.2
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    • pp.164-169
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    • 1995
  • An endogenous cryptic plasmid, pBL1, which has been used to construct plasmid vectors for coryneform bacteria producing amino acids, was eliminated from Brevibacterium lactofermentum. The pBL1 was partially digested with Sau3AI and the resulting DNA fragments were subcloned into a suicide vector pEM1 which contains a kanamycin-resistant (km$^{r}$) gene. KM$^{r}$ B. lactofermentum transconjugants were obtained by conjugal transfer of the pEM1 derivatives containing pBL1 DNA fragments from Escherichia coli into B. lactofermentum. A km$^{r}$ transconjugant was analyzed to contain a plasmid pEB14, which occurred in vivo by homologous recombination between pBL1 and the conjugal-transferred plasmid. The pEB14 including the pEM1-derived km$^{r}$ gene was found to be lost concomitantly with km$^{r}$ phenotype, resulting in the construction of a pBL1-free strain of B lactofermentum. Based on transformation efficiencies and plasmid stability, the resultant pBL1- free strain is more useful than wild strain as a host cell for genetic manipulation. It could be concluded that foreign plasmid DNAs are efficiently isolated and analyzed from the pBL1-free strain because of the absence of endogenous pBL1 plasmid.

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The Status of Laboratory Animal Production and Visions in the 21st Century - Review -

  • Gartner, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.7
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    • pp.1142-1151
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    • 1999
  • Today, laboratory animal production has decreased world-wide to half the number estimated in 1970 of more than 100 Mio. This is due to the cell-biological assays which replaced animal experimentation as a first allround method to solve biomedical problems. Animal experimentation remains the most significant experimental method for the study of higher organized physiological systems and their multifactorial connections. This requires maximal uniformity of all quantitative traits among the animals used for such studies (mainly mice and rats) and stability of these traits for reproducing such studies at any time world-wide. The success of the developed methods for the standardization of laboratory animals was analyzed and were found only partly be acceptable. Getting a higher degree of uniformity among standardized inbred animals is blocked by "intangible variance". This is caused by influences of ooplasm, shown by experimental twin and clone studies. Manipulation of this component of variance is essential in the future. - Genetic drifts impair the necessary stability of biological traits. There are a few disadvantages associated with the cryopreservation of embryos and other methods are required. - Dogs and cats were replaced by pigs as laboratory animals. A new line of animal production will evolve over the next 25 years with similarities to the present laboratory animal production, because in future pigs were used as donors for xenotransplants for men.

Efficacy Evaluation of Tissue Inhibitor of Metalloproteinases-2 and Endostatin on Angiogenesis (Tissue Inhibitor of Metalloproteinases-2와 Endostatin의 혈관신생 제어 효능 평가)

  • Kim, Soo-Hyeon;Cho, Young-Rak;Yoon, Hyun-Jae;Ko, Hee-Young;Kim, Pyeung-Hyeun;Seo, Dong-Wan
    • YAKHAK HOEJI
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    • v.54 no.6
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    • pp.488-493
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    • 2010
  • Therapeutic manipulation of angiogenesis, the formation of new vascular sprouts from existing capillaries, is one of the promising strategies for treatment of human diseases such as cancer, arthritis, and cardiovascular disorder. In the present study, we examined the effects and molecular mechanism of tissue inhibitor of metalloproteinases-2 (TIMP-2) and endostatin on fibroblast growth factor-2 (FGF-2)-stimulated endothelial cell proliferation, migration and adhesion in vitro, and angiogenesis in vivo. TIMP-2 and endostatin showed potent anti-angiogenic activity in vitro and in vivo. These effects appear to be mediated through different angiogenic signaling pathways. Collectively, our findings demonstrate that TIMP-2 and endostatin strongly inhibit FGF-2-induced angiogenic responses, and the establishment of fast and reproducible evaluation system in vitro and in vivo for the development of anti-angiogenic biomaterials and therapeutics.

Characterization of Microfluidic Channels using DVD Pick-up Fluorescent Scanner (광 픽업 방식 형광스캐너를 이용한 미소유체 특성 분석)

  • Yim, Vit;Kim, Jae-Hyun;Lee, Seung-Yop;Park, Jung-Yul
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.32 no.12
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    • pp.1102-1106
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    • 2008
  • Microfluidics deals with the behavior, precise control and manipulation of fluids at a micro scale. It has become increasingly prevalent in various applications such as biomedical applications (diagnostics, therapeutics, and cell/tissue engineering), inkjet head, and fuel cells etc. The issue of inspection and characterization of microfluidics has emerged as a major consideration in design, fabrication, and detection of microfluidic devices. In this paper, we characterize a diffusion based mixing in Y-microchannel using a fluorescent optical scanner based on a DVD pick-up module, which is widely used in optical storages. Using fluorescent dye, we measure the fluorescent intensity that represents the mixing patterns in Y-microchannel. We also compare these experimental results with computational fluid dynamics (CFD) simulation ones. It is shown that the proposed optical scanner can be used as an alternative measurement system with high performance and cost-effectiveness, compared to conventional optical tools such as epifluorescent microscopes using high resolution CCD camera and confocal microscopes with photomultiplier (PMT) detectors.

Production of Therapeutic Glycoproteins throgh the Engineering of Glycosylation Pathway in Yeast

  • Roy, Samir-Kumar;Yasunori Chiba;Yoshifumi Jigami
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.4
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    • pp.219-226
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    • 2000
  • The application of recombinant DNA technology to restructure metabolic net-work can change metabolite and protein products by altering the biosynthetic pathways in an organism. Although some success has been achieved, a more detailed and thorough investigation of this approach is certainly warranted since it is clear that such methods hold great potential based on the encouraging results obtained so far. In last decade, there have been tremendous advances in the field of glycobiology and the stage has been set for the biotechnological production of glycoproteins for therapeutic use. Today glycoproteins are one of the most important groups of pharmaceutical products. In this study the attempt was made to focus on identifying technologies that may have general application for modifying glycosylation pathway of the yeast cells in order to produce glycoproteins of therapeutic use. The carbohydrates of therapeutic recombinant glycoproteins play very important roles in determining their pharmacokinetic properties. A number of biological interactions and biological functions mediated by glycans are also being targeted for therapeutic manipulation in vivo. For a commercially viable production of therapeutic glycoproteins a metabolic engineering of a host cell is yet to be established.

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Manipulation of Antioxidative Mechanism in Chloroplasts

  • Kwon, Suk-Yoon;Lee, Haeng-Soon;Kwak, Sang-Soo
    • Proceedings of the Botanical Society of Korea Conference
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    • 1999.07a
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    • pp.79-84
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    • 1999
  • Oxidative stress is one of the major environmental stresses to plants. Reactive oxygen species (ROS) generated during metabolic processes damage cellular functions and consequently lead to cell death. Fortunately plants have in vivo defense system by which the ROS is scavenged by enzymes such as superoxide dismutase (SOD) and ascorbate peroxidase (APX). In attempts to understand the protection mechanism of plant against oxidative stress, we developed transgenic tobacco (Nicotiana tabacum cv. Xanthi) plansts thet expressed both SOD and APX in chloroplast using Agrobacterum-mediated transformation and evaluated their protection capabilities against methyl viologen (MV, paraquat) -mediated oxidative damage. Three double transformants (CAI, CA2, and CA3) expressed the chimeric CuZnSOD and chimeric APX in chloroplast, and one transformant (AM) expressed the chimeric APX and chimeric MnSOD in chloroplast. In addition, we obtained three lines of transformants (C/Al, C/A2, and A/C) that expressed the APX and SOD than control plants, and more resistant to oxidative stress caused by MV. TRansformants (C/A and A/C) overexpressing MnSOD, CuZnSOD and APX at the same time showed the highest resistance to MV-mediated oxidative stress among the transformants.

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표면특성이 제어된 기능성 나노 입자의 전자 및 의공학적 응용

  • 박영준;이준영;김중현
    • Proceedings of the Korea Crystallographic Association Conference
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    • 2002.11a
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    • pp.54-55
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    • 2002
  • The fabrication, characterization and manipulation of nanoparticle system brings together physics, chemistry, materials science and biology in an unprecedented way. Phenomena occurring in such systems are fundamental to the workings of electronic devices, but also to living organisms. The ability to fabricate the surface of nanoparticles Is essential in the further development of functional devices that incorporate nanoscale features. Even more essential is the ability to introduce a wide range of chemical and materials flexibility into these structures to build up more complex nanostructures that can ultimately rival biological nanosystems. In this respect, polymers are potentially ideal nanoscale building blocks because of their length scale, well-defined architecture, controlled synthesis, ease of processing and wide range of chemical functionality that can be incorporated. In this presentation, we will look at a number of promising polymer-based nanoparticle fabrication strategies that have been developed recently, with an emphasis on those techniques that incorporate nanostructured polymeric particles into electronic devices or biomedical applications. And functional nanoparticles deliberately designed using several powerful process methods and their application will be discussed. Nanostructured nanoparticles, what we called, implies dispersed colloids with the size ranged from several nanometers to hundreds of nanometer. They have extremely large surface area, thus it is very important to control the morphology or surface functionality fitted for adequate objectives and properties. Their properties should be controlled for various kind of bio-related technologies, such as immunomagnetic cell separation, drug delivery systems, labeling and identification of lymphocyte populations, extracorporeal and hemoperfusion systems, etc. Well-defined polymeric nanoparticles can be considered as smart bomb or MEMS.

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Auxin-responsive SMALL AUXIN UP RNA genes : recent research progress and its application for crop improvement (옥신 반응 SMALL AUXIN UP RNA 유전자의 최근 연구 동향 및 작물 개량을 위한 적용)

  • Lee, Sang Ho
    • Journal of Plant Biotechnology
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    • v.40 no.2
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    • pp.59-64
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    • 2013
  • Auxin is a key plant hormone which regulates overall plant growth development. A number of researches to investigate auxin signaling identified three major classes of early auxin response genes: AUX/IAA, GH3 and SMALL AUXIN UP RNA (SAUR). Among these genes, in planta functions of SAUR gene family are largely ambiguous, while both AUX/IAA and GH3 genes are analyzed to mediate negative feedback on auxin response. SAUR genes encode small plant-specific proteins. SAUR gene products are highly unstable and transiently expressed in the tissue- and developmental-specific manners in response to auxin and various environmental stimuli. In the decades, molecular and genetic approaches to elucidate in planta functions of SAURs have been hampered by several factors such as the unstable molecular features and functional redundancy among them. However, a series of recent studies focusing on several subgroups of SAUR gene family made significant progress in our understanding of its biochemical and physiological functions. These works suggest that many SAUR proteins mainly regulate auxin-related cell expansion and auxin transport. In this review, the recent progress in SAUR research and prospects for crop improvement through its genetic manipulation are discussed.

Enhanced Lycopene Production by UV-C Irradiation in Radiation-Resistant Deinococcus radiodurans R1

  • Kang, Chang Keun;Yang, Jung Eun;Park, Hae Woong;Choi, Yong Jun
    • Journal of Microbiology and Biotechnology
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    • v.30 no.12
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    • pp.1937-1943
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    • 2020
  • Although classical metabolic engineering strategies have succeeded in developing microbial strains capable of producing desired bioproducts, metabolic imbalance resulting from extensive genetic manipulation often leads to decreased productivity. Thus, abiotic strategies for improving microbial production performance can be an alternative to overcome drawbacks arising from intensive metabolic engineering. Herein, we report a promising abiotic method for enhancing lycopene production by UV-C irradiation using a radiation-resistant ΔcrtLm/crtB+dxs+ Deinococcus radiodurans R1 strain. First, the onset of UV irradiation was determined through analysis of the expression of 11 genes mainly involved in the carotenoid biosynthetic pathway in the ΔcrtLm/crtB+dxs+ D. radiodurans R1 strain. Second, the effects of different UV wavelengths (UV-A, UV-B, and UV-C) on lycopene production were investigated. UV-C irradiation induced the highest production, resulting in a 69.9% increase in lycopene content [64.2 ± 3.2 mg/g dry cell weight (DCW)]. Extended UV-C irradiation further enhanced lycopene content up to 73.9 ± 2.3 mg/g DCW, a 95.5% increase compared to production without UV-C irradiation (37.8 ± 0.7 mg/g DCW).