• Journal of Sericultural and Entomological Science
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• v.51 no.1
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• pp.73-77
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• 2013

Silkworm transgenesis scientists have done some genetic modification work on multivoltine silkworms, but that type of silkworms is less commercial feasible. They are easy to manipulate, because they breed all year round. But the commercial silkworm variety must undergo hydrochloric acid treatment at a high temperature to be artificially hatched. Hydrochloric acid penetrates through the holes in the silkworm eggs, fatally damaging their reproduction. So it had been thought that altering the properties of the commercial silkworm variety would be very difficult. So we have tried to make from diapause to non-diapause eggs using diapauses varieties, 'Backokjam' and 'Jam 124'. At present, our group has establishing the conditions for non-diapause eggs. Oviposited eggs after 40 ~ 60 hours were incubated for 24 hours at $15{\sim}20^{\circ}C$ with dark condition. Non-diapause eggs were completely induced. The hatching rate, molting rate and pupation rate of non-diapause 'Jam 124' and 'Backokjam' eggs showed no differences compared to diapause eggs. When transgenic silkworm using the non-diapause eggs, the hatching rate showed that non-diapause eggs induced from diapause were 40 ~ 70%, diapause eggs treated with artificial incubation were 10 ~ 30%, and polyvoltine strains, HM eggs were 30 ~ 50%. Therefore, we suggest that modification techniques of the commercial silkworm eggs adequate for silkworm transgenesis can be used to develop transgenic silkworms more easily.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.93-93
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• 2003

소의 수정란이식에 있어서 성공적인 임신을 위해서는 수란우의 적절한 영양상태, 양질의 수정란, 이식 시술자의 기술력과 수란우와 수정란의 적절한 동기화가 필수 요인이라 사료된다. 특히, 적절한 동기화를 위해서 발정관찰은 필수적이지만 번거롭고 장시간동안 관찰을 해야 하는 단점이 있다. 따라서 본 연구에서는 이러한 단점을 보완할 수 있도록 발정동기화 처리방법을 비교, 검토하여 수정란이식에 있어서 효과적인 발정동기화 방법을 모색하고자 실시하였다. 발정동기화 방법은 Fig 1과 같이 CIDR처리방법(A군)과 GnRH 처리방법(B군)을 사용하였으며, 황체의 등급은 직장검사와 초음파 진단기를 이용하여 직경이 2cm 이상, 1~2cm와 1cm 미만의 황체를 각각 1등급, 2등급과 3등급으로 분류하였다. 또한 본 실험에 공시된 수정란은 체외 생산된 한우 배반포기의 수정란을 사용하였으며, 1등급의 황체를 가진 수란우만을 선별하여 비외과적인 방법으로 황체가 존재하는 자궁각심부에 이식하였다. 임신진단은 이식 후 45~60일에 직장검사와 초음파진단을 이용하여 실시하였다. 발정동기화처리결과는 Table 1에서 보는 바와 같이 A군과 B군에서 발정발현율이 각각 100%와 96%로 나타났으며, 이식하기에 적합한 1등급 황체의 출현율이 65.4%와 56%로 A군에서 다소 높게 나타났다. 발정동기화 처리방법에 따른 수정란 이식 후 수태율은 A군에서 신선란과 동결란일 때 각각 66.7%와 60%로 나타나 B군의 22.2%와 0%의 결과보다 유의적으로 높은 결과를 나타내었다. 따라서 본 연구의 결과로 미루어 볼 때 수정란 이식을 위한 발정동기화 방법은 CIDR 처리방법을 적용하는 것이 GnRH 처리방법 보다 효율적이라 사료된다.다. 특히 기능황체에서의 특이적 발현 spot을 확인할 수 있었다. 이러한 결과들로부터 황체의 progesterone분비기능의 역할을 수행하기 위한 단백질들이 전, 중기에 발현된다는 것을 알 수 있고 퇴행황체에서는 발현이 안되고 있는 것을 알 수 있다. 이러한 결과를 토대로 다른 양상을 띤 spot을 분리하여 어떤 단백질인지를 분석하여 각각의 황체단백질의 특성을 규명할 수 있을 것으로 사료된다.적율(HCT)을 이루고 있음을 알 수 있었다. 적혈구 평균용적(Mean Corpuscular Volume ; MCV), 평균적혈구혈색소량(Mean Corpuscular Hemoglobin ; MCH), 평균적혈구혈색소농도(Mean Corpuscular Hemoglobin Concentration ; MCHC) 그리고 혈소판(Platelets) 분석결과 형질전환돼지가 일반돼지보다 약간 높은 수치를 나타냈으며 변화양상 또한 유사한 결과를 나타내었다. 이와 같은 시험분석결과를 토대로 사람 조혈촉진유전자(hEPO)가 형질전환된 돼지는 유즙으로 발현할 수 있도록 형질전환 되었음에도 불구하고 헤모글로빈 및 적혈구가 증가함으로서 형질전환돼지 개체의 혈장으로도 사람 조혈촉진인자가 분비하고 있음을 간접적으로 알 수 있었다. 정상돼지 보다 형질전환돼지가 약 30% 높은HCT 수준을 보였으며 이러한 현상은 사람에서는 적혈구증다증(erythrocytosis)으로 분류되고 있다. 이에 대한 고찰은 형질전환돼지 자체의 생리적 문제점(side effects)에 대한 해결과 더불어 기존의 인간질병에 대한 모델동물로서의 이용 가능성을 제시할 수 있는 것으로 사료된다.mu\textrm{m}$ 300BG는 56.32$\mu\tex

• Korean Journal of Poultry Science
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• v.32 no.4
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• pp.225-229
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• 2005

Microinjection of recombinant retrovirus beneath the blastoderm of non-incubated chicken embryo is now the most widespread method for generating transgenic chickens, but transgenesis rates are very low. So to improve this problem, we first introduced retrovirus vector carrying RSV-GFP gene to an one-cell embryo culture system. To investigate whether retrovirus could work on an one-cell chicken embryo, we microinjected the concentrated retrovirus stocks into the germinal disc of one cell or stage-X chicken embryos. Analysis of reporter gene expression on day 4 embryos showed that GFP expression was observed in the only stage-X chicken embryo but was not in the one-cell embryo group. These results suggest that retrovirus system is the most efficient method to generate transgenic chickens in the stage-X embryo.

• Proceedings of the Korea Society of Information Technology Applications Conference
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• 2006.06a
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• pp.185-207
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• 2006

산업단지는 지난 30년간 한국 산업의 성장을 이끌어 온 발전모형으로서 존재하여 왔으나, 최근 지식에 기초한 혁신창출형 경제체제가 국가 및 지역사회의 경쟁력을 위한 핵심요소로 부각되면서 '효율성' 측면에서 그 의미가 크게 퇴색되었다. 이를 위한 대안으로서 '클러스터'가 대두되어 다양한 분석연구가 수행되고 있으며, 정부와 지방자치단체들은 이를 바탕으로 각자의 특색에 맞는 클러스터 조성 정책을 펼치고 있다. 그러나, 기존의 연구들은 클러스터의 종류 및 발전단계에 관한 프레임워크 제시 등의 이론적 수준에 국한되어 있거나, 지역사례 연구를 통한 성공요인분석(CFS) 및 단순한 정책방향 제시 수준에 머물러 있는 한계를 보이고 있다. 본 연구는 '클러스터'에 관한 선행연구를 분석해 보고, 클러스터의 중요한 판단기준이 되는 군집도와 네트워크 연계 정도를 기준으로 한 '$2{\times}2$ 클러스터 전략격자모형'을 효과적인 클러스터 구축전략 수립을 위한 이론적 틀로서 제시하였다. 또한, 분석틀에 실질적인 사례로서 '충북지역의 SW산업'을 전략격자모형에 대응시켜 분석함으로써 전략격자의 유용성을 제시하였다. 이를 위해, 충북지역의 SW 공급업체와 수요업체를 대상으로 설문조사를 실시, 분석한 후 그 결과를 전략격자모형에 대응시켰다. 그 결과, 충북지역의 SW산업은 아직 산업단지 수준에 있는 것으로 분석되었고 충북의 SW산업의 충북 내의 수요만으로는 더 큰 성장이 어려운 것으로 분석, 지역 내에서의 수요창출을 목표로 하는 '단일 클러스터' 구축보다는 지역적 제약을 벗어난 '매가 클러스터'의 구축으로 지역 내외에서의 수요창출이 가능한 클러스터의 구축을 그 대안으로 제시하였다.${\alpha}$에 E. coli Jm109의 plasmid pBX19, pBR322를 전이시켰다. 6. L. lactis ssp. lactis 균주에 lysozyme 처리시 30${\sim}$80%의 생존율을 보였으며, 대부분의 L. acidophilus 균주의 경우 약 70%의 생존율을 보였다. L. casei 102S의 경우는 45분간 처리 시에도 100%의 생존율을 보였다. 8. L. lactis ssp. lactis 균주에 pLZ12를 6.0kV에서 전이시킨 결과 12.5kV에서보다 형질전환 효율이 훨씬 높았으며 lysozyme 처리에 의해 형질전환 효율이 증가되었다. 9. L. acidophilus 균주에 pLZ12를 전이시 6.0kV에서는 전이가 모두 이루어졌으나, 12.5kV에서는 L. acidophilus WIESBY와 NCFM에서 전이가 이루어지지 않았으며, lysozyme 처리 후 pLZ12를 전이시켰을 때 12kV보다 6.0kV에서 형질전환 효율이 증가되었다. 10. Gene Pulser와 Progenitor II를 사용하여 pLZ12를 L. lactis ssp. lactis 균주에 전이하였을 때 Gene Pulser에 비해 Progenitor II의 형질전환 효율이 현저히 떨어졌다. L. acidophilus HY7008과 HY7001은 두 기기 모두 형질전환이 이루어졌으나, L. acidophilus WEISBY와 NCFM은 Progeni-tor II에서 전이가 일어나지 않았으며, Gene Pulser에서 전이균주를 얻어 두 electroporator간에 형질전환 효율의 차이를 보였다. 11. L. casei 102S에 pLZ12

• Journal of Ginseng Research
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• v.27 no.1
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• pp.37-42
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• 2003

For study about introduce of gene connected with disease and transformation system of gingseng, chitinase gene cloned from soybene and disease resistant gene were carried out for expression and transformation of plant using Agrobacterium. The disease resistance gene(DR-49), 35S-35S-AMV, has been constructed. The disease resistance gene and chitinase gene were introduced into the binary vector pRD 400, which were mobilized into Agrobacterium tumefaciens faciens strain MP 90 and LBA 4404 harboring disarmed Ti-plasmid. As a result of induce transformants using ginseng embryo and petiole, multi shoots were formed on MS medium supplemented 1 mg/ι 2,4-D and 0.5 mg/ι kinetin. Also transformation by cotyledonwas effective on MS medium supplemented 1 mg/ι 2,4-D and 0.5 mg/ι kinetin, transformation percent of disease resistant gene and chitinase gene were showed 18%, 14% respectively. As transformed tissue is under pre-embryoid condition, normal shoot is required through the process of matured embryo.

• Journal of The Korean Society of Grassland and Forage Science
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• v.26 no.2
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• pp.77-82
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• 2006

An efficient transformation system for the production of transgenic plants has been developed for tall fescue (Festuca arundinacea Schreb.) via Agrobacterium-mediated transformation of seed-derived callus. From the point of morphogenetic capacity, three types of callus were selected. High frequency of plant regeneration was obtained by selection of type II callus, and the plant regeneration frequency was 52.6% when embryogenic callus were cultured on the regeneration medium. Supplementation of the media with 10 mg/L $AgNO_3$ and 40 mg/L cysteine enhanced frequencies of plant regeneration up to 65.3%. The highest transformation efficiency was also obtained when type II callus were inoculated with Agrobacterium. Southern blot analysis of PCR products of transgenic plants demonstrated that transgenes were successfully integrated into the genome of tall fescue. Efficient regeneration system and transformation established in this study will be useful for molecular breeding of tall fescue through genetic transformation.

• Microbiology and Biotechnology Letters
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• v.14 no.3
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• pp.213-218
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• 1986

Hybrid plasmid pEA24, shuttle vector YRp7 carrying amylase gene of Bacillus amyloliquefaciens, was transformed to yeast Saccharomyces cerevisiae, and the expression of B. amyloliquefaciens amylase gene in yeast was investigated. The frequency of transformation to S. cerevisiae DBY747 with YRp7 was increased by treatment of 40% polyethylene glycol (MW 4, 000), PH 7.0, at 3$0^{\circ}C$, and by regeneration used 2% top agar. The amount of cellular amylase activity produced by S. cerevisiae containing pEA24 was 2% of that secreted from B. amyloliquefaciens, but in case of S. cerevisiae transformant, the amylase secreted was not detected. A comparison of genetic stability of pEA24 and YRp7 plasmids in yeast was carried out by cultivation of transformants in tryptophan-supplement-medium. The pEA24 plasmid was more unstable than YRp7 in S. cerevisiae. The size of pEA24 extracted from S. cerevisiae transformants was found to be identical with that from E. coli transformants by agarose gel electrophoresis.

• FLOWER RESEARCH JOURNAL
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• v.18 no.1
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• pp.1-8
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• 2010

Sensitivities of PLBs of four Phalaenopsis cultivars, P. 'Taisuco Windian', P. 'Nancy Amour', P. 'Pink Twilight' and P. 'Taipei Gold' to kanamycin, spectinomycin and hygromycin at different concentrations (0, 25, 50, 100, 200, and $400mg{\cdot}L^{-1}$) were examined. Hygromycin was favorable for selecting the transformants in the genetic transformation of Phalaenopsis as PLBs of four cultivars were all dead at even $25mg{\cdot}L^{-1}$ hygromycin. Responses of PLBs of P. 'Maki Watanabe' and P. 'Brother Lawrence' to DL-phosphinothricin (PPT) were determined at different concentrations (0, 0.1. 0.25, 0.5, 1.0, 2.0, 2.5, and $5.0mg{\cdot}L^{-1}$) and $0.5mg{\cdot}L^{-1}$ PPT was thought to be suitable for selecting the transformants of Phalaenopsis. The optimum conditions for Agrobacterium cocultivation with Phalaenopsis PLBs were examined using a two-step cocultivation method in Dtps. 'City Girl' and A. tumefaciens LBA4404. In the first infection period in a 1 : 10 suspension of Agrobacterium to a VW medium, 1 hr infection showed the highest PLB survival ratio. And then, PLBs were cocultivated with a bacterial strain and a 3-day cocultivation period was better for Phalaenopsis PLBs than a prolonged period. Agrobacterium tumefaciens strains LBA4404 (pTOK233) and EHA105 (pGA643) were used to compare their efficiency on the genetic transformation of Phalaenopsis PLBs. The PLBs infected with EHA105 survived more than those infected with LBA4404 after two days in a dark condition and two weeks in light condition on a selective medium. About 1,000 PLBs for each of P. 'Maki Watanabe' and P. 'Brother Lawrence', and each bacterial strain of AGL1 (pCAMBIA3301) and LBA4404 (pTOK233) were used for the regeneration of transgenic plants. The bacterial strain AGL1 had a higher genetic transformation efficiency than LBA4404, with no significant difference between cultivars. In this study, 11 hygromycin-resistant plantlets and 32 PPT-resistant plantlets were produced, but these putative transgenic plantlets need further examinations.

• Korean Journal of Plant Tissue Culture
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• v.24 no.5
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• pp.269-272
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• 1997

Cotyledonary petioles of Brassica napus cocultivated with Agrobacterium vectors for 72 h were transferred to MS medium with 0.5 mg/L NAA, 2.0 mg/L BA, 30 mg/L kanamycin, 100 mg/L cefotaxime, 30 g/L sucrose, 3 mg/L $\textrm{AgNO}_{3}$ and 2 g/L Gelrite. The cotyledonary petioles with green shoots were selected at a frequency of 17.5% in a selection medium and then rooted. Southern blot analysis confirmed the rolC and NPT IIgenes were incorporated into the regenerated plants. The stable inheritance of rolC gene was confirmed in progeny test of transgenic plants.

• Korean Journal of Plant Tissue Culture
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• v.24 no.1
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• pp.37-41
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• 1997

The in vitro regeneration and genetic transformation systems in hot pepper(Capsicum annuum L.) have not been routinely available, which has been a major limiting factor in the application of new genetic manipulations. An efficient procedure to regenerate whole pepper plants and to generate transgenic plants expressing a foreign gene was established. A relatively high frequency of plant regeneration was observed when hypocotyl and cotyledon explants were cultured on MS medium supplemented with NAA 0.1 mg/L plus zeatin 2.0 mg/L or IBA 10.0 mg/L plus BAP 1.0 mg/L. Addition of AgNO$_3$5 $\mu$M to these media improved the regeneration frequency up to 8%. For plant transformation, hypocotyl and cotyledon explants of hot pepper were precultured on shoot induction media without kanamycin added for 2 days, and then cocultured with Agrobacterium tumefaciens pDY183 for 2 days. Putative transformants were obtained from selection media containing 100 mg/L kanamycin sulfate and 500 mg/L carbenicillin. Putatively selected transformants were confirmed by amplification of selectable marker genes (ADA and NPT II) by polymerase chain reacion. Successful transcripts of ADA gene were detected by Northern blot analysis. Enzyme activity of ADA was also examined by spectrophotometric analysis, and expression of ADA gene in hot pepper suggests the potential application of ADA gene as a selectable marker in plants.