• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.60-60
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• 2003

소의 난포란과 난구세포의 체외배양시 plasminogen activators(PAs)의 생산을 보고하였다 (Choi 등, 1998). 따라서 본 연구는 돼지 난포란 및 난구세포의 체외성숙시 PAs의 생산을 SDS-PAGE와 Zymogram을 이용하여 조사하였다. PCOCs는 도축암퇘지의 난소로부터 채취하여, 난구세포가 충실한 것만 선별하였으며, 실험구에 사용될 난구세포는 pipetting에 의해 분리하여 이용하였다. 돼지 신선정액은 D-PBS로 1,500 rpm, 5분간 2회 원심분리하여 정장물질을 제거하고, 3회째는 5mM caffein이 함유된 BO(Brackett과 Oliphant, 1985) 배양액으로 세정하였다. 처리한 돼지 정액은 1$\times$$10^{8}$ cells/$m\ell$로 조정하여 20${\mu}\ell$씩 분주하고 0, 1, 2, 3 또는 4시간 동안 39$^{\circ}C$ 5% $CO_2$, 95% 공기인 배양기에서 수정능획득을 유도하였다. 배양이 완료된 정액은 20${\mu}\ell$의 sample buffer(5% SDS, 20% glycerol, 0.0025% bromophenol blue 그리고 0.125M Tris HC1 buffer)에 넣어 -7$0^{\circ}C$ 동결기에 보관하였다. 전기영동은 4% stacking gel과 10% separating gel로 분리하였으며, 20 mA에서 90분간 실시하였다. Zymogram은 Choi 등(1988)의 방법에 따라 실시하여 PAs의 생산을 확인하였으며, 이상의 실험은 3반복을 실시하였다. 시험구 전체에서 urokinase type plasminogen activator(uPA)가 확인되었으며, 체외수정능 획득시간에는 차이가 없었다. 두 종류의 고분자량의 uPA의존성 영역이 나타났으며, 분자량은 65kD과 62 kD이었다. 이러한 결과로 볼 때 Hart 등(1986)이 uPA의 경우 다양한 영역의 분자량 변이를 확인할 수 있었다고 한 것과 동일하였으며, 돼지 정자가 체외수정능 획득시 uPA를 생산하는 것을 확인할 수 있었다.

• KSBB Journal
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• v.15 no.6
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• pp.573-577
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• 2000

The optimum fermentation conditions for the production of cellulose by a newly isolated Acetobacter sp. A9 were determined in static cultures. The strain was able to produce cellulose at $25-30^{\circ}C$ with a maximum at $30^{\circ}C$. Cellulose production occurred at pH 6.5-8.0 with a maximum at pH 6.5. The optimal culture medium was found to consists of 1.0% glucose, 1.0% yeast extract, 0.7% polypeptone, 0.15% acetic acid and 0.02% succinic acid. Cellulose production by Acetobacter sp. A9 followed the growth curve. Highest cellulose production, under optimum conditions, was $24.1m^2$, although this strain typically produced only $12.1 g/m^2$ in the basic medium. Cellulose production also depended on the depth and volume of the medium.

• Journal of the Korean Applied Science and Technology
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• v.37 no.6
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• pp.1698-1705
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• 2020

Bacteria were collected from the thumb surface of the twenty young adults that are 20 to 25 years old and cultured on the Luria-Bertani agar. The 16S rDNA of the cultured bacteria was amplified by polymerase chain reaction(PCR) and DNA sequence of the PCR products analyzed. Total 14 different bacterial species were identified by comparing their 16S rDNA sequence with the data in genbank. It appears that each individual has 2.5 different bacterial species in average. Staphylococcal species were the most abundant among the identified bacteria and Micrococcus luteus was the second. Staphylococcal species were isolated at similar frequency between male and female donors but Micrococcus luteus was isolated more frequently from female than male donors. The result obtained in this study might be useful in research of dermatic diseases, searching for new drugs for those diseases and development of new cosmetics.

• Journal of Embryo Transfer
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• v.22 no.2
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• pp.137-141
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• 2007

Antioxidants were well known to be essential supplements in the complex media and serve as a reservoir of oxygen. In this study, Hanwoo COCs (cumulus oocytes complexes) were matured and developed in L-cysteine-TCM199 and analyzed metaphase chromosome. Maturation rate of Hanwoo COCs were 73.4%, 94.6% in 0.1% PVA, 0.1 mM L-cysteine, respectively and showed significantly different between the treatments (p<0.05). Blastocyst formation were revealed 20.3%, 10.0% in 5% FBS+TCM199, 0.1 mM L-cysteine+1% BSA, respectively. There were no significant difference among treatment groups. Metaphase chromosome were showed 18.3%, 12.0% in 5% FBS-TCM199, 0.1 mM L-cysteine, respectively and analyzable chromosome were 6.1%, 4.0% and had no differences between the treated groups. In the case of in vitro developmental stages, metaphase chromosome were showed 18.3%, 12.0% in $4{\sim}16$ cells stage, 43.1%, 13.0% in morulae stage and 94.8%, 100.0% in blastocyst stage. These results suggested L-cysteine has beneficial role for in virto maturation and development in Hanwoo COCs.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.86-86
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• 2002

면양과 염소가 최근 수십년동안 세계여러 나라에서 번식생리의 연구를 위한 모델로 사용되어 왔는데, 체내수정란의 생산에 관한 영역도 유럽을 중심으로 활발하게 연구되어왔다. 수정란생산을 위한 발정동기화방법, 과배란처리 및 수정란회수방법 기술은 현재 상당히 많은 기술진척이 이루어진 상태이나, 우리나라 고유의 재래유전자원인 흑염소에는 이를 위한 기술이 미진한 실정이므로 본 실험에서는 흑염소의 체내수정란생산기술을 확립하여 재래가축 유전자원보존을 위한 기초기술을 마련하고자 한다. 축산기술연구소 남원지소에서 사육하고 있는 체중 20kg 이상의 건강한 흑염소를 이용하여 발정동기화를 위해 controlled intravaginal drug release(CIDR)를 질내에 14일 동안 삽입하고, 과배란처리는 FSH를 CIDR 삽입 12, 13, 14일째에 12시간 간격으로 점감법으로 총20mg을 투여하였으며, PGF$_2$a를 13일째 FSH와 함께 투여하였다. CIDR는 14일째의 아침에 제거하였다. 수컷과의 교미는 CIDR제거 24시간후에 GnRH를 투여와 동시에 실시하였으며, 채란은 교미후 3일째에 외과적인 방법으로 실시하였다. CIDR처리경과에 따른 progesterone농도는 CIDR 주입시 바로 수치가 상승하여 제거전까지 6~12ng/m1의 농도를 유지하였으며, 제거즉시 2ng/ml 이하로 떨어졌다. 채란시 평균 배란점은 16.5개, 미배란난포 9.8개였으며, 회수수정란은 6.0개를 나타내어 채란율은 36.4%를 나타내었다. 회수된 수정란의 발달단계는 4-cell 78.9%, 2-cell 5.3%, fragmentation 15.8%를 나타내었다. 이와 같은 체내수정란생산방법을 기반으로 하여 이후 수정란의 동결 및 수정란이식기법에 관한 연구를 수행한다면 우리나라의 재래가축인 흑염소의 유전자원 장기보존과 생산성향상에 기여할 것으로 사료된다.배양액에 30 embryos/50ul 소적으로하여 38.8$^{\circ}C$, 5% $CO_2$의 탄산가스 배양기에서 각각 7일간 배양을 실시하였다. 조사된 결과는 SAS/STAT를 이용하여 통계분석을 실시하였다. 체외수정 12시간 후에 난자 급속 염색법으로 염색을 실시한 결과, 모든 처리구에서 핵성숙률(76.4~95.2%), 정자침투율(51.1~66.9%), 웅성전핵형성률(95.2~100%), 다정자침입률(18.2~25.6%) 및 평균침입정자수(1.2~l.4개)에서 유의적인 차이가 인정되지 않았다. 체외배양 48시간 난할률을 조사한 결과, 처리구별 차이(53.9~67.9%)는 인정되지 않았으나, 배양 7일째 배반포형성률은 각각 14.5, 25.4, 17.3 및 12.4%로서 25uM의 $\beta$-ME처리구가 유의적(P<0.05)으로 높은 배발달률을 나타내었고, 총세포수에 있어서는 대조구와 처리구간 유의적인 차이가 인정되지 않았다. 따라서 돼지 난포란을 성숙배양할 때, 25uM $\beta$-ME를 첨가배양하는 것이 양질의 돼지체외수정란을 생산하는 하나의 방법으로 조사되었다.다.natural objects and was popular at the time of Yukjo Dynasty, and there are some documents of that period left both in Japan and Korea. "Hyojedo" in Korea is supposed to have been influenced by the letter design. Asite- is also considered to have been "Japanese Letter Jobcheso." Therefore, the purpose of this study is to look into the origin

• Horticultural Science & Technology
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• v.18 no.6
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• pp.802-807
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• 2000

This study was conducted to propagate the arbuscular mycorrhizal fungi in vitro using the hairy root of carrot transformed by Agrobacterium rhizogenes with Ri t-DNA. Mycorrhizal spores and roots in sudangrass plants were wet-sieved, surface-sterilized and inoculated onto the hairy root of carrot on the Modified Strullu & Romand (MSR) medium. The mycorrhizal spores of Glomus sp. propagated in vitro for 12 weeks was about $50{\mu}m$, and the shapes of spores were round or elliptic. Spores were formed mainly at the middle of the hyphae. Number of mycorrhizal spores propagated using dual culture of the transformed carrot roots and the mycorrhizal inoculum for 12 weeks were about 1,200 per plates.

• Journal of Bio-Environment Control
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• v.13 no.2
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• pp.112-119
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• 2004

Objective of this research was to evaluate the effects of nutrient solution strength and Arbuscular Mycorrhizal Fungi (AMF, Glomus sp.) on growth and flowering of potted miniature rose (Rosa hybrids L. cv 'Scarlet'). To achieve this, plants cultured with six different strength of Japanese Horticultural Experiment Station solution (0.125, 0.25, 0.5, 1.0, 2.0, and $4.0\;{\times}\;{full}$ strength) and inoculated with AMP at cutting and transplanting. Leachate EC increased as solution strength were elevated. The leachate EC were not different between non-inoculated plants and AMF treatment at cutting, but significantly decreased when plants were inoculated with AMF at transplanting. The elevated strength of nutrient solution resulted in decrease of leachate pH. When plants were inoculated AMF at transplanting, leachate pH was lower than those of non-inoculated plants and inoculated with AMF at cutting. At harvesting (93 days after transplanting), plant height, leaf width, number of branches and shoot fresh and dry weight of rose 'Scarlet' increased with elevated nutrient solution strength. AMF treatment at transplanting of potted rose 'Scarlet' showed the best results in growth such as chlorophyll content, number of flowers, and shortening the days required to flower. The content of N, P, K, and Mn in leaf tissue of potted rose increased by elevated nutrient solution strength and AMF treatment, while the tissue Na contents decreased by an AMF treatment.

• Journal of Korean Society of Forest Science
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• v.77 no.2
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• pp.208-215
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• 1988

The leaf mesophyll protoplasts of Populus alba ${\times}$ glandulosa were isolated from leaf of plantlet in vitro and cultured for plant regeneration. The MS medium (minus $NH_4NO_3$) with 0.5 mg/l BAP and 2.0 mg/l 2, 4-D showed the moderate frequency of dividing protoplasts cultured by the liquid plating method during the first week of culture. The percentage of colony formation was revealed the highest frequency by the gauze contained semi-solid agar plating method after 5 weeks cultured. Ridding out the gauze, the micro-callus was formed on the same semi-solid medium in 8 weeks after protoplasts culture. For proliferation of callus, mini-callus was transferred on the MS solid medium with 0.5 mg/l 2, 4-D and 0.1 mg/l BAP 12 weeks after culture. Shoot regeneration occurred when the calli derived from protoplasts were cultured on MS medium with 1.0 mg/l zeatin and such shoots could be readily rooted on the one half strengthen MS medium with non-phytohormone. Rooting shoots were planted in green-house 22 weeks after protoplast culture.

• Journal of Plant Biotechnology
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• v.32 no.3
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• pp.217-223
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• 2005

Effects of growth regulators on growth of adventitious roots and accumulation of steroidal saponins, such as dioscin, prosapogenin A and prosapogenin C, in cultures of Dioscorea nipponica were determined. The maximum growth of adventitious roots was observed in MS medium supplemented with 30 g/L sucrose and 1.0 mg/L NAA. Addition of BA in combination with NAA appeared to be no effective in the growth of adventitious roots. Among the twenty different adventitious roots formed from different seeds, strain No. 10 was selected based on production ability of dioscin, and its stability through the successive liquid culture. During the first 4 weeks of incubation, contents of steroidal saponins in adventitious roots were negligible but the contents were markedly increased at 5 weeks of incubation. Dioscin and prosapogenin C content in IBA-treated adventitious roots were significantly higher than those in NAA-treated roots. However, content of prosapogenin A was not significantly different among NAA or IBA level. Results provide that liquid culture of adventitious roots of D. nipponica have a potential for mass production of dioscin including prosapogenin A and prosapogenin C.

• Korean Journal of Plant Tissue Culture
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• v.26 no.2
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• pp.121-126
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• 1999

Calli were induced from the petiole explants of Pimpinella brachycarpa on MS medium supplemented with 0.5 mg/L 2,4-D and 0.1 mg/L BA after four weeks of culture. Compact clusters of small and dense cells among these calli were selected and suspension-cultured as the source of embryogenic calli. When transferred to MS medium with 0.1 mg/L NAA, the suspension-cultured cells grew to embryogenic callus. Somatic embryos derived from these embryogenic calli developed into plantlets. The cDNA library was constructed in the embryogenic callus and in order to screen the cDNA library, these cDNAs were plated at a density 1.5 $\times$ 10^5 plaques per 15 cm petridish. Among 19 clones showing preferential hybridization with petiole HD-Zip gene, five clones were obtained after second screening. Four clones among them, were highly homologous to P. brachycarpa shoot-tip Phz4 gene, but one clone, Phc5 was about 1.5 kb which has an extra 163 bp to 5' upstream of Phz4. The Phc5 was 1,531 bp containing poly A tails of 18 bases. ATG start codon for Phc5, was located at position 284 with an open reading frame of 906 by which encodes a polypeptide of 302 amino acids. The Phc5 protein revealed that the polypeptides between 135 and 195 contain a homeodomain as the `leucine zipper' motif.