To product the blue fluorescent protein (AmCyan) expressed cocoon, we were fused AmCyan cDNA to the heavy chain gene and injected the gene into a silkworm. AmCyan was one of the existing violet fluorochromes and originally derived from the fluorescent protein amFP486. AmFP486 was cloned from the sea anemone Anemonia majano (GenBank accession number AF168421), and belongs to the family of fluorescent proteins (FPs) isolated from coral reef organisms. The AmCyan fusion protein, each with N- and C- terminal sequences or the fibroin H-chain, were designed to be secreted into the lumen of the posterior silk glands. The expression of the AmCyan/H-chain fusion gene was regulated by the fibroin H-chain promoter. The use of the 3xP3 EGFP as a marker allowed us to rapidly distinguish transgenic silkworm. Mixtures of the donor and helper vectors were micro-injected into 300 eggs of bivoltine silkworms (Baegokjam). EGFP fluorescence was observed in 3 broods of transgenic silkworms under a florescence stereomicroscope. The cocoon was displayed strong blue fluorescence, proving that the fusion protein was present in the cocoon. Accordingly, we suggest that the AmCyan gene expressed cocoon will be enable the production of the novel biomaterials based on the transgenic silk.

High luminance LED fluorescent light is replaced by halogen light in recent light parts. The housing cover of LED fluorescent light is used many several materials. Though deformity character has less value, polycarbonate cover has cheap, high strength and it is used general purpose. This study is several purposes. 1. No entering foreign matters during continuous extrusion manufacturing process in cutting process, 2. No deformity, exact cutting of pc length and finding method of cutting manufacturing process for polycarbonate cover

폐쇄형 묘생산 시스템에서 청색LED, 적색LED, 백색형광등을 인공광원으로 이용한 가운데 파프리카의 육묘시 생장 특성과 정식 후 생장 및 초기 수량을 분석하고자 본 연구가 수행되었다. 폐쇄형 시스템에서 파프리카 육묘용 환경조건은 광주기 16/8h, 평균 PPF 204μmol·m-2·s-1, 기온 26/20℃, 상대습도 70%이었다. 육묘 후 21일째에 백색형광등과 LED 하에서 생장된 파프리카 묘의 엽장, 엽폭, 엽면적 등 잎 관련지표뿐만 아니라 지상부 생체중과 건물중, 엽록소함량 등이 자연광 처리구에 비해서 크게 나타났다. 청색 LED, 적색LED 및 자연광 처리구에서의 엽면적은 대조구인 형광등 처리구와 비교할 때 각각 63%, 63%, 28%에 해당하였다. 또한 청색LED, 적색LED 및 자연광 처리구의 지상부 건물중은 각각 대조구의 64%, 50%, 22%로 나타났다. 정식 후 18일째에 엽수는 대조구에서 44매로 가장 크게 나타났다. 적색LED, 청색LED 및 자연광 처리구의 엽수는 대조구에 비해서 각각 86%, 81%, 48%로서 정식 시기와 비교할 때 엽수의 차이가 줄어들었다. 정식 후 114일째에 초장은 청색LED와 적색LED 처리구에서 상대적으로 작게 나타났다. 이러한 결과는 단색LED 하에서 육묘된 파프리카의 줄기 신장이 정식 후에 억제된 것으로 판단된다. 초기 4주 동안 수확된 파프리카는 청색LED 3.5개/plant, 적색LED 3.3개/plant, 자연광 1.0개/plant으로서 대조구 2.2개/plant에 비해서 각각 159%, 150%, 45%로 나타났다. 초기수량은 적색LED 453g/plant, 청색LED 403g/plant, 자연광 101g/plant으로서 대조구 273g/plant와 비교할 때 각각 166%, 148%, 37%로 나타났다. 한편 적색LED 처리구에서의 평균 중량은 136g으로서 다른 처리구와 비교할 때 상대적으로 큰 과실이 생산되었다. 한편 정식 후 온실에서의 재배기간이 길어짐에 따라 인공광 처리구와 자연광 처리구에서 수량 차이가 없었다. 이러한 결과를 종합하면 LED 또는 형광등을 인공광원으로 이용한 조건에서 육묘된 파프리카의 정식 후 초기 생육이 양호하였으며, 초기 수확이 자연광 처리구에 비해서 1주 정도 빠르게 이루어졌음을 알 수 있다. 따라서 LED 또는 형광등과 같은 인공광원이 파프리카 육묘에 이용될 경우 묘소질의 향상, 조기 수확 및 초기 수량의 증대가 기대된다.

Random amplified polymorphic DNA (RAPD) and fluorescent amplified fragment length polymorphism (FAFLP) analyses were executed on a total of 28 Salmonella spp., including 6 ATCC reference strains, 2 isolates from outbreaks of food poisoning in Gwangju, and 20 isolates from carcasses. For RAPD analysis, four primers, named P1254, 23L, OPA-4, OPB-17 were used producing amplification fragments ranged from 0.18kb to 2.6kb. As a result, 5 types using P1254, 5 types using 23L, 3 types using OPA-4, and 6 types using OPB-17 and a total of 18 RAPD types were achieved. For FAFLP analysis, bacterial genomic DNA was digested with endonucleases EcoRⅠ and MseⅠ, site-specific adaptors were ligated, and PCR amplification was carried out with an EcoR1 adaptor-specific primer labelled with fluorescent dye. Amplified fragments, which were separated on a polyacrylamide sequencing gel ranged from 35bp to 300bp were analysed. Results were displayed as a dendrogram with genetic distance. Twenty two Salmonella isolates and 6 reference strains were divided into 14 groups in a level of 0.136 genetic distance. In conclusion, Salmonella isolates of chicken carcasses have different genetic properties when compared to reference strains and isolates from outbreak of food poisoning.

In this study, the entrapped number is investigated on the UV light with different illuminance to fluorescent bait cage for swimming crab in order to find the appropriate illuminance which has the best attraction effect of fluorescent bait cage for pots. In addition, preference to the light, arrival time and residence time at light area are compared and analyzed to fluorescent bait cage and non-fluorescent bait cage for American lobster at the UV light and ordinary light according to the illuminance condition. Pot with red non-fluorescent bait cage at the no lighting (<0.01lux), pot with blue fluorescent bait cage at the 20W UV lighting (0.16lux) and pot with blue fluorescent bait cage at the 30W UV lighting (0.22lux) were soaked for 6 hours and the entrapped number of swimming crab was examined. The mean entrapped number of swimming crab in pot with red non-fluorescent bait cage at the no lighting (<0.01lux) was 1.0, but the mean entrapped number of swimming crab in pot with blue fluorescent bait cages at the 20W UV lighting (0.16lux) and 30W UV lighting (0.22lux) were 1.4 and 0.4, respectively (P<0.05). The rate of preference to the blue fluorescent bait cage at the UV lighting shows 1.6-4.8 times higher than that of preference to the red non-fluorescent bait cage at the ordinary lighting. In addition, The rate of preference to the blue fluorescent bait cage at the UV lighting is higher when the illuminance of ordinary light is same as or is lower than that of UV light (P<0.05). However, the preference to the light depending on gender shows no significant difference (P>0.05). The arrival time to UV light area of lobster is shown as 1.2-2.4 times faster than that to ordinary light area. Generally, it is shown that arrival time to UV light area is faster than the arrival time to ordinary light area when the illuminance of ordinary light is the same as or lower than that of UV light (P<0.05). However, arrival time to the light area depending on gender shows no significant difference (P>0.05). The residence time at UV light area of lobster is 1.2-1.7 times longer than that at ordinary light area. The residence time depending on different illuminance of ordinary light and genders showed no significant difference (P>0.05).

수처리 분리막 공정에서 막 오염 제어 기술은 현장 적용 기술 및 경제성 확보에 있어 매우 중요하다. 본 연구에서는 형광 나노 입자 및 형광 분광 분석법을 도입함으로써 수처리 분리막 공정에서 막 오염 정도를 실시간으로 측정 모니터링 할 수 있는 센싱 기술을 개발하고자 하였다. 막 오염 정도를 모니터링 할 수 있는 분리막 제조를 위해 세 종류의 형광물질 OB, FP, KCB를 담지한 다공성 polysulfone (PSf) 비대칭 막을 제조하였다. 형광 분광 분석 시스템을 이용하여 분리막 표면에서의 오염 정도를 실시간으로 측정한 결과, 형광 물질을 첨가한 막은 막 오염이 진행됨에 따라 형광 신호가 크게 감소함을 보여 막 표면 오염층의 모니터링 분석이 가능함을 확인하였다.

Many methods have been developed for more efficient gene delivery and expression in human cells. A number of studies have been performed in achieving successful gene delivery and expression conditions. We investigated differential gene expression patterns after delivery adenoviral vector containing green fluorescent protein(GFP) gene into human cancer cell lines. We constructed recombinant adenoviral Ad-CMV-GFP containing CMV promoter and GFP gene. The efficiency of gene expression was assessed by observation GFP expressing cells using fluorescent microscopy after transfer of Ad-CMV-GFP in concentrations of 0.1μl. 1μl. 10μl. At first, we evaluated expression patterns of gene in several human cancer cell lines, gastric adenocarcinoma cell line AGS was showed high level of GFP expression compared with colorectal adenocarcinoma cell line HT-29. After transfer 0.1μl of Ad-CMV-GFP in AGS, we could found that GFP expression cells were observed in next day and highly increased 2 days. While, small number of GFP expressing cells were examined in HT-29 and SNU-C4. Therefore, these data showed that AGS was expressed the highest level of GFP and almost AGS cells seems to express GFP in concentration of 1μl of Ad-CMV-GFP. GFP expression pattern in HT-29 reveal that expression was low in next day after gene transfer but significantly increase expression level in 2 days. In case of SNU-C4, GFP expression increased with increasing concentration of Ad-CMV-GFP and t ransfer times. For examine effects of transfer times in small amount gene, we transfer in concentration of 0.1μl Ad-CMV-GFP and detected GFP expression patterns after 2 days or 4 days. As a result, expression level of GFP in AGS was increase about 2 fold after 4 days compared with 2 days, but any difference of GFP expression levels were not showed in HT-29 and SNU-C4. Our study suggested that adenovirus was very efficient gene transfer vector for gene expression in human cancer cell lines. In addition to, we also demonstrated that gene expression patterns was dependent on each human cell lines. Therefore, further studies will be needed to confirm the optimum conditions for efficient gene delivery and expression in each target cell lines with consideration to cellular properties.

Treponema denticola is a gram-negative anaerobe that can cause periodontal disease. The adhesion of this bacterium to host tissues is considered to be the primary event in the colonization and infection of a host. Fibrinogen is generally found in damaged tissues resulting from periodontitis. The binding ability of T. denticola to fibrinogen may therefore be an important virulence factor in inducing periodontal diseases. It has been reported recently that oral spirochetes can be labeled with fluorescent fatty acids and we speculated that this labeling method could be used in an oral spirochete binding assay. The binding of several different strains of T. denticola to immobilized human fibrinogen was therefore tested using the fluorescent fatty acid labeling method. In the case of immobilized fibrinogen, the T. denticola ATCC 35405 strain showed saturable binding to immobilized fibrinogen. Indeed, all four different T. denticola strains tested in this experiment, T. denticola ATCC 35405, T. denticola ATCC 33520, T. denticola ATCC 35404 and T. denticola OTK showed binding to fibrinogen. The fluorescent fatty acid labeling method thus shows utility in binding assays for T. denticola, different strains of which can generally bind to immobilized fibrinogen.

In order to develop the substitutive materials for natural baits of swimming crab pots, the fluorescent characteristics of the baits were analyzed, and the preference of fluorescent dyes were investigated by the mean entrapped catch number to the pots through the water tank experiments and fishing experiments. On the investigation of fluorescent characteristics by the 5 kinds of baits, mackerel, krill, manila clam, pig's fat and chicken's head which were used in substitutive baits for test in the UV long wave(365nm) area, it showed clear blue fluorescence in the skin of mackerel, shell of krill, manila clam and bill of chicken's head, and green fluorescence in the mackerel s muscle and internals, and yellow fluorescence in the pig's fat and chicken's head. On the investigation of fluorescent characteristics by the bait cages in the UV short wave(254nm) and long wave(365nm) area, it showed each green, red and blue fluorescence in the cylinderical or hexahedral red plastic bait cages which were painted each green, red and blue fluorescence dyes, but it showed yellowish green flourescence in the cylinderical or hexahedral red plastic bait cage which was painted yellow fluorescent dye. On the preference investigation of the fluorescent dyes of swimming crabs by the 5 kinds of the bait cages which were put the mackerel in the non-fluorescent red plastic cage(RFN), red, yellow, green and blue fluorescent plastic cages(RF, YF, GF, BF) each, nonfluorescent red plastic cage(RFN) was entrapped mean 2.0(6.7%), but blue fluorescent plastic cage(BF) was mean 5.0(16.7%) and it was more 2.5 times comparing to RFN, and red fluorescent cage(RF) was same level and green fluorescent cage(GF) was 50% of catch number comparing to RFN, and yellow fluorescent cage(YF) was entrapped nothing(F 46.324, P 〈 0.05). On the investigation of the entrapped catch number to the pots which were put the mackerel in the blue fluorescent hexahedral plastic cage(HP) and blue fluorescent silicon mackerel model cage(SM), HP was mean 3.4(11.3%) and it was a little more comparing to SM which was entrapped mean 3.2(10.7%)(t 0.775, P 〉 0.05). Fishing experiments on the preference investigation of swimming crabs by the pots which were put in the non-fluorescent red plastic cage(RFN) and blue fluorescent plastic cage(BF) were conducted 3 times. Mean catching number and weight of RFN were 71.7 ind.(18.3%) and 16.9kg(64.3%), and those of BF were 93.0 ind.(23.1%) and 19.8kg(64.5%), respectively.

Recent studies on nuclear transfer and induced pluripotent stem cells have demonstrated that differentiated somatic cells can be returned to the undifferentiated state by reversing their developmental process. These epigenetically reprogrammed somatic cells may again be differentiated into various cell types, and used for cell replacement therapies through autologous transplantation to treat many degenerative diseases. To date, however, reprogramming of somatic cells into undifferentiated cells has been extremely inefficient. Hence, reliable markers to identify the event of reprogramming would assist effective selection of reprogrammed cells. In this study, a transgene construct encoding enhanced green fluorescent protein (EGFP) under the regulation of human Oct4 promoter was developed as a reporter for the reprogramming of somatic cells. Microinjection of the transgene construct into pronuclei of fertilized mouse eggs resulted in the emission of green fluorescence, suggesting that the undifferentiated cytoplasmic environment provided by fertilized eggs induces the expression of EGFP. Next, the transgene construct was introduced into human embryonic fibroblasts, and the nuclei from these cells were transferred into enucleated porcine oocytes. Along with their in vitro development, nuclear transfer embryos emitted green fluorescence, suggesting the reprogramming of donor nuclei in nuclear transfer embryos. The results of the present study demonstrate that expression of the transgene under the regulation of human Oct4 promoter coincides with epigenetic reprogramming, and may be used as a convenient marker that non-invasively reflects reprogramming of somatic cells.

Oral spirochetes are anaerobes known as one of causative agents for periodontal diseases. In this study, we investigated the possibility of utilizing fluorescent fatty acids for labeling oral spirochetes. Bacterial labeling was standardized with three different lengths of fluorescent fatty acids: 5-octadecanoylaminfluorescein (OAF), 5-dodecanoylamin-fluorescein (DAF), and 5-hexadecanoylaminfluorescein (HAF). Among these fatty acids, OAF showed the best labeling activity. Treponema denticola ATCC 35405 was totally saturated to the maximum when incubated with OAF 1μM/mℓ for 1 hour. Treponema vincentii LA-1 also increased in fluorescence in proportion to incubation time length and the concentration. In conclusion, these findings showed the possibility that the fluorescent fatty acid can be used for labeling oral spirochetes.