• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2006년도 The 6th Intermational Symposium on Developmental Biotechnology
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• pp.10-10
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• 2006

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2006년도 The 6th Intermational Symposium on Developmental Biotechnology
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• pp.49-49
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• 2006

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2002년도 추계학술대회
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• pp.109-115
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• 2002

Gene therapy is to treat and cure diseases by an introduction of therapeutic genes in defective cells or tissues of human body. Gene delivery system, gene expression system, and therapeutic gene are three core elements for gene therapy. The efficient delivery of therapeutic genes and appropriate gene expression are the crucial issues for therapeutic outcome of gene delivery. Because it can be used in common for the treatment and cure of various diseases, gene delivery system is the most important core element for a successful gene therapy. Viruses are naturally evolved to transfer their genomes into host cells efficiently. This ability has made vectorologists exploit viruses as attractive vehicles for the delivery of therapeutic genes. Viral vectors based on adenovirus (Ad) and adeno-associated virus (AAV) have been often used for gene delivery in laboratory. Ad and AAV vectors derived from human DNA viruses differ greatly in their life cycle, expression level and duration of transgenes, immunogenicity, and vector preparation. Both vectors can be used as effective tools for gene therapy and more recently in functional genomics. Here, the characteristics of Ad and AAV vectors are discussed.

• 미생물학회지
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• 제52권3호
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• pp.298-305
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• 2016

본 연구에서는 p-cresol 초기 분해에 관여하는 기존의 lap과 pcu 유전자군 외에 새로운 pch 유전자군을 Pseudomonas alkylphenolica KL28로 부터 동정하였다. 이 유전자군(pchACXF-pcaHG-orf4-pcaBC)은 p-cresol을 ${\beta}$-carboxy-cis,cis-muconate로의 전환을 촉매할 수 있는 효소를 암호화하는 것을 알 수 있었다. 이 유전자 군은 영국에서 분리된 Pseudomonas putida NCIMB 9866과 9869의 plasmid에서 유래된 pch 유전자 군과 동일하여, 이들 유전자군은 종간 horizontal gene transfer로 전달되었을 가능성을 제시하였다. 각 유전자군의 관련 유전자의 변이와 gfp 레포터를 갖는 프로모터의 발현 분석을 통해 3개의 분해 유전자군이 모두 p-cresol의 분해에 관여하는 것을 알 수 있었으며, pch 유전자는 p-cresol에 의해 유도되며, 고체 및 액체 배지에서도 pcu 유전자군이 가장 높게 발현되는 것을 확인할 수 있었다. 또한 pcu 유전자 변이주는 p-cresol을 이용하여 버섯모양의 공중체(aerial structure) 형성하지 않았으므로, 탄소원의 이용 속도가 다세포 구조 형성에 영향을 주는 중요한 요소 중의 하나임을 알 수 있었다.

• 한국수정란이식학회지
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• 제17권2호
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• pp.101-108
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• 2002

Human Prourokinase (proUK) offers potential as a novel agent with improved fibrin specificity and, as such, may offer advantages as an attractive alternative to urokinase that is associated with clinical benefits in patients with acute peripheral arterial occlusion. For production of transgenic cow as human proUK bioreacotor, we conducted this study to establish efficient production system for bovine transgenic embryos by somatic cell nuclear transfer (NT) using human prourokinase gene transfected donor cell. An expression plasmid for human prourokinase was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker gene, and human prourokinase target gene into a pcDNA3 plasmid. Cumulus cells were used as donor cell and transfected with the expression plasmid using the Fugene 6 as a carrier. To increase the efficiency for the production of transgenic NT, development rates were compared between non-transfected and transfected cell in experiment 1, and in experiment 2, development rates were compared according to level of GFP expression in donor cells. In experiment 1, development rates of non-transgenic NT embryos were significantly higher than transgenic NT embryos (43.3 vs. 28.4％). In experiment 2, there were no significant differences in fusion rates (85.4 vs. 78.9％) and cleavage rates (78.7 vs. 84.4％) between low and high expressed cells. However, development rates to blastocyst were higher in low expressed cells (17.0 vs. 33.3％), and GFP expression rates in blastocyst were higher in high expressed cells (75.0 vs. 43.3％), significantly.

• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 2003년도 제3회 국제심포지움 및 학술대회
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• pp.109-109
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• 2003

Even though success in birth of live offspring from nuclear transfer(NT) using somatic cells in many species, detailed information on processes or mechanisms of development are not well known. Cytoplasm of bovine oocyte has been known to support the development of nuclear transferred embryos using nuclear donor cells from different species. Therefore, interspecies NT might be used to find answers of some questions in basic aspect of nuclear transfer In this study, we examined the developmental potential of reconstructed embryos when bovine oocyte as a cytoplasm recipient and mouse embryonic fibroblast as a nuclear donor were used. The nuclear transfer units were aliocated in Group 1 (murine block media and normal media) and Group 2. (bovine block media and normal media). NT units were not blocked at 2-cell stage regardless of types of medium. On mouse media, poor development of interspecies NT units was observed compared to bovine media. However, as NT units cultured in bovine normal medium, embryos developed over 8-cell stage. Further studies performed to increase the developmental rate in condition of antioxidant treatment. Despite low development, bovine-murine interspecies nuclear transferred embryos could develop to blastocysts and they showed that blastocyts rate of antioxidant group was superior to those of non-antioxidant group. Next, we investigated gene expression pattern which is carried out for zygotic activation. The Xist gene is expressed in female mouse embryo after zygotic activation of 4-cell stage. But interspecies nuclear transferred embryos do not express Xist gene at 4-cell stage. As a result, it is suggested that the bovine cytoplasm controls the early preimplantation development in interspecies NT However, the development of later stages might require genomic control from transferred donor nucleus. Therefore, even though the involvement of several other factors such as mitochondrial incompatibility, effective development of embryos produced by interspecies NT requires proper genomic activation of donor nucleus after overcoming the cytoplasmic control of recipient oocytes.

• Fisheries and Aquatic Sciences
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• 제9권2호
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• pp.97-100
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• 2006

To determine whether the tetracycline resistance genes tet (34), tet (M), and tet (S) can be transferred among bacteria, we used a filter mating experiment allowing intimate cell-cell contact between donor and recipient. The tet(34) gene, conveyed on a chromosome of Vibrio species (No. 6 and SW-42) was not transferred to Escherichia coli JM109, suggesting that it is not transferred among bacterial species. The tet (M) gene was transferred from three Vibrio strains (4-E, SW-18, and SW-38) to E. coli at frequencies of $8.5{\times}10^{-5}\;to\;2.1{\times}10^{-6}$. The tet(S) gene was transferred from Lactococcus garvieae KHS98032 to E. coli at a frequency of $1.8{\times}10^{-6}$. Transconjugated recipients showed increased minimum inhibitory concentrations against oxytetracycline. Although the donors possess the Tn916-Tn1545 transposons, they were not detected in transformed recipients, suggesting that the transfer of tet(M) and tet(S) is mediated by elements or mechanisms. Two ribosomal protect protein genes were also transmissible from marine bacteria to E. coli, suggesting gene hopping among marine, terrestrial, and human environments.

• International Journal of Industrial Entomology and Biomaterials
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• 제3권1호
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• pp.75-81
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• 2001

We have constructed a novel recombinant Bombyx mori nuclear polyhedrosis virus (BmNPV) producing the green fluorescent polyhedra. For the production of the fluorescent polyhedra, partial polyhedrin gene containing KRKK as nuclear localization site from the BmNPV polyhedrin gene and the green fluorescent protein (gfp) gene were introduced under the control of p10 promoter of BmNPV. The recombinant BmNPV was stably produced fluorescent polyhedra in the infected Bm5 cells and the morphology of the fluorescent polyhedra was similar to that of wild-type BmNPV. The fluorescent polyhedra had 32 kDa native polyhedrin and 41 kDa fusion protein. From these data, we have further developed a novel BmNPV p10-based transfer vector producing recombinant polyhedra with foreign gene Product. The novel BmNPV P10-based transfer vector is composed of partial polyhedrin gene, factor Xa, and multiple cloning sites.

• IMMUNE NETWORK
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• 제7권4호
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• pp.179-185
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• 2007

Background: Adenovirus (Ad) vectors have been widely used for many gene therapy applications because of their high transduction ability and broad tropism. However, their utility for cancer gene therapy is limited by their poor transduction into cancer cells lacking the primary receptor, coxsackievirus and adenovirus receptor (CAR). Methods: To achieve CAR-independent gene transfer via Ad, we pretreated Ad with 1,2-dioleoyl-3-trimethylammonium propane (DOTAP) and analyzed their transduction efficiency into cancer cells in vitro and in vivo comparing with the virus alone. Results: Treatment of DOTAP significantly increased adenoviral gene transfer in tumor cells in vitro. Moreover, DOTAP at an optimum dose $(10{\mu}g/ml)$ enhanced IL-12 transgene expression by fivefold in tumor, and twofold in serum after intratumoral injection of adenovirus expressing IL-12N220L (Ad/IL-12N220L). In addition, cotreatment of DOTAP decreased tumor growth rate in the Ad/IL-12N220L-transduced tumor model, finally leading to enhanced survival rate. Conclusion: Our results strongly suggest that DOTAP could be of great utility for improving adenovirus-mediated cancer gene therapy.

• Toxicological Research
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• 제17권
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• pp.173-183
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• 2001

Although there are still many technical difficulties to be overcome, recent advances in the molecular and cellular biology of gene transfer have made it likely that gene therapy will soon start to play an increasing role in clinical practice. However. safety issues are raised from vector system. It is not clear whether it is safe to incorporate genes into nuclear DNA. Little is known about the antigenicity of gene product which the immune system is encountering. In this review, some safety-related topics are introduced and discussed.