• Title/Summary/Keyword: selection and concentration

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Introduction and Expression of PAP gene using Agrobacterium in Scrophularia buergeriana Miquel (Agrobacterium을 이용한 PAP 유전자의 현삼으로 도입 및 형질발현)

  • Yu, Chang-Yeon;Seong, Eun-Soo;Lim, Jung-Dae;Huang, Shan-Ai;Chae, Young-Am
    • Korean Journal of Medicinal Crop Science
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    • v.9 no.2
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    • pp.156-165
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    • 2001
  • Exogeneous application of pokeweed antiviral protein (PAP), a ribosomal-inacivating protein in the cell wall of Phytolacca americana (pokeweed) protects heterologous plants from viral and fungal infection. A cDNA clone of PAP introduced into Scrophularia buergeriana Miquel by thransformation with Agrobacterium tumefaciences. For plant transformation, explants were precultured on shoot induction medium without kanamycin for 2-5 day, and then they were cocultured with Agrobacterium for 10 minutes. The explants were placed on co culture medium in dark condition, $28^{\circ}C$ for 2days. After explants were washed in MS liquid medium, they were transferred into selection medium including kanamycin 50mg/L (MS salts+1mg/ l BAP+2mg/ l TDZ+0,2mg/ l NAA+MS vitamin+3% sucrose+0.8% agar, pH5.8). From PCR analysis, NPT II band was confirmed in transgenic plant genome and showed resistance against fungi in antifungal activity test. Micro assay to which protein extracted from transgenic line were added, revealed hyphae growth inhibition and no spore germination at high concentration. The characteristics of inhibited hyphae was represented transparent and thin. Expression of PAP in transgenic plants offers the possibility of developing resistance to viral and fungal infection.

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Bioinformatic Analysis of the Canine Genes Related to Phenotypes for the Working Dogs (특수 목적견으로서의 품성 및 능력 관련 유전자들에 관한 생물정보학적 분석)

  • Kwon, Yun-Jeong;Eo, Jungwoo;Choi, Bong-Hwan;Choi, Yuri;Gim, Jeong-An;Kim, Dahee;Kim, Tae-Hun;Seong, Hwan-Hoo;Kim, Heui-Soo
    • Journal of Life Science
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    • v.23 no.11
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    • pp.1325-1335
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    • 2013
  • Working dogs, such as rescue dogs, military watch dogs, guide dogs, and search dogs, are selected by in-training examination of desired traits, including concentration, possessiveness, and boldness. In recent years, genetic information has been considered to be an important factor for the outstanding abilities of working dogs. To characterize the molecular features of the canine genes related to phenotypes for working dogs, we investigated the 24 previously reported genes (AR, BDNF, DAT, DBH, DGCR2, DRD4, MAOA, MAOB, SLC6A4, TH, TPH2, IFT88, KCNA3, TBR2, TRKB, ACE, GNB1, MSTN, PLCL1, SLC25A22, WFIKKN2, APOE, GRIN2B, and PIK3CG) that were categorized to personality, olfactory sense, and athletic/learning ability. We analyzed the chromosomal location, gene-gene interactions, Gene Ontology, and expression patterns of these genes using bioinformatic tools. In addition, variable numbers of tandem repeat (VNTR) or microsatellite (MS) polymorphism in the AR, MAOA, MAOB, TH, DAT, DBH, and DRD4 genes were reviewed. Taken together, we suggest that the genetic background of the canine genes associated with various working dog behaviors and skill performance attributes could be used for proper selection of superior working dogs.

Transformation of Populus nigra × P. maximowiczii Using Agrobacterium tumefaciens vectors (Agrobacterium tumefaciens vector를 이용(利用)한 양황철의 형질전환(形質轉換))

  • Son, Suk Gyu;Hyu, Jung Oh
    • Journal of Korean Society of Forest Science
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    • v.87 no.2
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    • pp.164-172
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    • 1998
  • This study was conducted to find the optimum transformation condition using Agrobacterium harboring promoterless GUS gene. The optimal medium for shoot induction from leaves of Populus nigra${\times}$P. maximowiczii was MS medium supplemented with $0.1mg/{\ell}$ NAA, $0.5mg/{\ell}$ BAP(94% regeneration frequency and 11.5 average number of shoot) According to the test using pBI121, the concentration of antibiotics for selection marker gene was $100mg/{\ell}$ kanamycin or $60mg/{\ell}$ geneticin in the SIM(shoot inducing medium) 3. Two weeks later, callus was induced in the SIM 3 and this callus grew up to 0.5-1cm shoots after 6 weeks in the new SIM 3. And the treatment with methylation inhibitor(5-azacytidine) led to a dramatic increase in foreign gene expression rate from 5.7% to 26.7%. The vector systems showed. different transformation efficiencies based on the fluorometric and histochemical GUS assay. In this study the vector systems used for transformation seemed to affect transformation frequency, in which pEHA101 yielded more transformants(35.9%) than LBA4404/pBI121 did(5.7%). This result indicated that pEHA101 was effective to insert the promoterless foreign gene into a poplar genome.

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Preselection of Bovine Blastocysts Expressing Exogeneous Gene Following Microinjection (외래유전자를 주입한 소 수정란에서 형질전환가능 수정란의 선발)

  • 공일근
    • Korean Journal of Animal Reproduction
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    • v.21 no.2
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    • pp.167-176
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    • 1997
  • This study was carried out to evaluate the potential for preselection of transgenic embryos prior to transfer into recipient animals. In these experiments, I used a 3.2 kb transgene which contained the neomycin resistance gene (neo) and lac Z gene driven by the $\beta$ actin promoter (iacZ Ineo). Oocytes were aspirated from abattoir ovaries, matured in TCM-199 supplemented with 10% fetal bovine serum (FBS), 5 ${\mu}\textrm{g}$/ml LH, 0.5 ${\mu}\textrm{g}$/ml FSH, 100 unit/ml penicillin, and 100 ${\mu}\textrm{g}$/ml streptomycin for 22 to 24 hrs then inseminated with a sperm suspension of 1 X 10$^6$ sperm/ml containing 5 ${\mu}\textrm{g}$/ml of heparin. At 18-20 hrs after insemination, cumulus cells were removed by vortexing and pronuclei of centrifuged zygotes microinjected with the lacZ/neo construct (3 ng/$\mu$l). All cultures were carried out in CR1aa with transfected BRL monolayers containing 3 mg/ml BSA, 20 $\mu$/ml NEM amino acids and 40 $\mu$I/ml BME amino acids. To identify the appropriate concentration of G418 for selection, non-microinjected zygotes were cultured in the presence of 0, 50, 100 and 200 $\mu$g/ml of G418. After 8 days of culture in these treatments, 44/145 (30.3%), 13/150 (8. 7%), 1/151 (0.7%) and 0/134 (0.0%) devel-oped to the blastocyst stage in 0, 50, 100 and 200 $\mu$g/ml of G418, respectively. A total of 1,127 zygotes were microinjected and placed into culture (without G418) and subsequently 710 (63.0%) cleaved. At 48 hrs post-injection, embryos ($\geq$2-cell) were randomly assigned to control (medium alone) or G418 (100 ${\mu}\textrm{g}$/ml) treatments. A control culture of 740 non-microinjected embryos from the same replicates of embryos were also placed into control medium. After 8 days in culture, 54/343 (15.7%) and 22/367 (6.0 %) of the microinjected embryos developed to the blastocyst stage in control and G418 media, respectively. A total of 151/740 (27.2%) of the non-microinjected embryos placed in the control medium developed to the blastocyst stage. The blastocysts in the control treatment had a mean of 70.7 ${\pm}$ 4.7 cells of which 23.1 ${\pm}$ 2.6 (32.7%) stained for $\beta$-Gal activity. B1astocysts in the G418 treatment had a mean of 48.8${\pm}$7.5 cells of which 40.3 ${\pm}$ 4.1 (82.6%) stained for $\beta$-Gal ac tivity (P<0.05). In the control treatment 26 of 30 (87.0%) blastocysts had some cells with $\beta$-Gal activity while all of the blastocysts in the G418 treatment had some cell with $\beta$-Gal ac tivity (P<0.05). However, ICM colonies in either control or G418 treatments were not expressed any epiblast cell except of trophetoderm celIs. The $\beta$-actin promoter/lacZ gene may not be e expression or silence expression in epiblast cells These results clearly show an enrichment of blastocysts expressing the transgene in the majority of their cells after culture in the presence of G418. The exogeneous gene was not express a and silence in ICM colonies especiallly epiblast cells except of trophectederm cells. Even though the higher rate cell number expressed of exogeneous gene in the G418 treatments, a total cell number was decrease significantly (P<0.05) of which might be also drop of the establishment of ES like-cell colonies and production of transgenic animals. However, futher studies need to determine the viability of these selected embryos and the avability of production of transgenic offspring.

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The Characteristics and Performances of Manufacturing SMEs that Utilize Public Information Support Infrastructure (공공 정보지원 인프라 활용한 제조 중소기업의 특징과 성과에 관한 연구)

  • Kim, Keun-Hwan;Kwon, Taehoon;Jun, Seung-pyo
    • Journal of Intelligence and Information Systems
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    • v.25 no.4
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    • pp.1-33
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    • 2019
  • The small and medium sized enterprises (hereinafter SMEs) are already at a competitive disadvantaged when compared to large companies with more abundant resources. Manufacturing SMEs not only need a lot of information needed for new product development for sustainable growth and survival, but also seek networking to overcome the limitations of resources, but they are faced with limitations due to their size limitations. In a new era in which connectivity increases the complexity and uncertainty of the business environment, SMEs are increasingly urged to find information and solve networking problems. In order to solve these problems, the government funded research institutes plays an important role and duty to solve the information asymmetry problem of SMEs. The purpose of this study is to identify the differentiating characteristics of SMEs that utilize the public information support infrastructure provided by SMEs to enhance the innovation capacity of SMEs, and how they contribute to corporate performance. We argue that we need an infrastructure for providing information support to SMEs as part of this effort to strengthen of the role of government funded institutions; in this study, we specifically identify the target of such a policy and furthermore empirically demonstrate the effects of such policy-based efforts. Our goal is to help establish the strategies for building the information supporting infrastructure. To achieve this purpose, we first classified the characteristics of SMEs that have been found to utilize the information supporting infrastructure provided by government funded institutions. This allows us to verify whether selection bias appears in the analyzed group, which helps us clarify the interpretative limits of our study results. Next, we performed mediator and moderator effect analysis for multiple variables to analyze the process through which the use of information supporting infrastructure led to an improvement in external networking capabilities and resulted in enhancing product competitiveness. This analysis helps identify the key factors we should focus on when offering indirect support to SMEs through the information supporting infrastructure, which in turn helps us more efficiently manage research related to SME supporting policies implemented by government funded institutions. The results of this study showed the following. First, SMEs that used the information supporting infrastructure were found to have a significant difference in size in comparison to domestic R&D SMEs, but on the other hand, there was no significant difference in the cluster analysis that considered various variables. Based on these findings, we confirmed that SMEs that use the information supporting infrastructure are superior in size, and had a relatively higher distribution of companies that transact to a greater degree with large companies, when compared to the SMEs composing the general group of SMEs. Also, we found that companies that already receive support from the information infrastructure have a high concentration of companies that need collaboration with government funded institution. Secondly, among the SMEs that use the information supporting infrastructure, we found that increasing external networking capabilities contributed to enhancing product competitiveness, and while this was no the effect of direct assistance, we also found that indirect contributions were made by increasing the open marketing capabilities: in other words, this was the result of an indirect-only mediator effect. Also, the number of times the company received additional support in this process through mentoring related to information utilization was found to have a mediated moderator effect on improving external networking capabilities and in turn strengthening product competitiveness. The results of this study provide several insights that will help establish policies. KISTI's information support infrastructure may lead to the conclusion that marketing is already well underway, but it intentionally supports groups that enable to achieve good performance. As a result, the government should provide clear priorities whether to support the companies in the underdevelopment or to aid better performance. Through our research, we have identified how public information infrastructure contributes to product competitiveness. Here, we can draw some policy implications. First, the public information support infrastructure should have the capability to enhance the ability to interact with or to find the expert that provides required information. Second, if the utilization of public information support (online) infrastructure is effective, it is not necessary to continuously provide informational mentoring, which is a parallel offline support. Rather, offline support such as mentoring should be used as an appropriate device for abnormal symptom monitoring. Third, it is required that SMEs should improve their ability to utilize, because the effect of enhancing networking capacity through public information support infrastructure and enhancing product competitiveness through such infrastructure appears in most types of companies rather than in specific SMEs.