• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.32-32
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• 2003

소 난자의 체외성숙 과정에서 세포질 내 단백질의 생산과 축적의 변화는 핵 및 세포질 성숙과 밀접한 관계가 있다는 보고가 있지만, 난자의 성숙과 관련된 특정 단백질의 종류에 대한 보고는 없었다. 따라서 본 연구는 한우 난자의 체외성숙과 관련된 단백질의 생산 및 축적의 변화와 그 종류에 대해서 검토하였다. 체외성숙 시간(4.5, 9, 13.5, 18 및 24시간)에 따른 배양액 내의 단백질 합성의 변화는 2D gel electrophoresis를 이용하였고, 단백질 spot에 대해서는 peptide mass fingerprinting(PMF) 방법을 이용하였다. 또한 단백질 측정 시간에 신선 체외성숙 배양액으로 교환 후 난포란의 핵성숙과 배발달율을 검토하였다. 그 결과 한우 난포란의 체외성숙 시간에 따라 배양액에서 단백질의 양 및 질적인 변화를 확인하였다. 그리고 총 296개 단백질 spot들을 확인하였고, 그 중 30개 spot에서 유의적인 변화가 인정되었다. 또한 유의적인 변화를 보인 spot에 대한 PMF 분석을 통하여 Apolipoprotein A-1 precursor, Alpha enolase, Aldose reductase, 43kDa collectin precursor, Heat shock 27kDa protein, Plasminogen activator inhibitor-1 precursor, Thrombospondin 1 및 Transitional endoplasmic reticulum ATPase가 동정되었다 그리고 총 단백질 합성 경향은 0∼4.5 시간에는 감소하였고, 13.5∼18시간에 증가 한 후 다시 감소하는 경향을 나타내었으며, 단백질의 종류도 시간대별로 현저한 변화가 있었다. 한편 단백질을 측정하는 시기에 신선 체외성숙 배양액으로 교환한 후 난포란의 핵성숙 및 배발달율을 검토한 결과 18시간 체외성숙군에서 9시간째의 교환이 유의적으로 높은 핵성숙을 나타내었으나, 배발달율에서는 유의성이 인정되지 않았다. 그러나 24시간 체외성숙군에서 18시간째의 배양액 교환은 8세포기 및 배반포 발달율이 유의적(P<0.05)으로 높았다. 연구 결과로부터 소 난자의 체외성숙 시간에 따른 단백질 합성 경향의 차이를 확인하였고, 유의적인 변화를 나타낸 8가지의 단백질을 분리할 수 있었으며, 향후 이들 작용기전에 대한 연구가 필요하다고 사료된다.

• Korean Journal of Plant Resources
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• v.18 no.2
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• pp.351-358
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• 2005

The effects of thiamine-HCl or polyvinylpyrrolidone (PVP) on growth and catechin contents of calli from Camellia sinensis leaves were investigated to improve catechin productivity in callus cultures on the selective medium. The growth of calli was great on the proliferation medium (the MS medium with 0.2 mg/L 2,4-D and 1.0 mg/L TDZ) supplemented with $20{\sim}30\;mg/L$ thiamine-HCl. Although the caffeine content was low in calli compared to young leaves, the total content of catechins was high in calli. Particularly,(-)-epicatechin (EC) which not detected in young loaves was also detected in calli on the propagation medium containing thiamine-HCl or PVP. In addition, the (-)-epicatechingallate (ECG) content was significantly higher in calli than in young leaves. In conclusion, the proliferation medium supplemented with 30 mg/L thiamine-HCl as a vitamin source seemed to be optimal condition for the growth and catechin production in callus culture.

• Journal of Bio-Environment Control
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• v.9 no.2
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• pp.79-84
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• 2000

The growth and quality differences between common thyme(Thymus vulgaris L.) and lemon thym (Thymus$\times$citriodorus) were investigated. They were grown in a hydroponics culture system (DFT) with a nutrient solution developed by the European Vegetable R&D Center in Belgium. Nutrient solution ionic strength used were 1.2, 2.4, 4.8 and 7.2 mS.cm-1. The dry matter of two thymes were increased with increasing ionic strength, while shoot length, root length and fresh weight were decreased. Chlorophyll content was higher in lemon thyme, while vitamin C content and essential oil content were higher in common thyme. The essential oil content showed a similar trend as those of growth. The main compostion of essential oil were thymol and carvacrol in common thyme, and geraniol and $\alpha$-citral in lemon thyme. The highest content of these compounds was obtained in 1.2mS.cm-1 treatment. Consequently, the ionic strength of 1.2mS.cm-1 was suitable for the production of quality thymes.

• Journal of the Korean Society of Food Science and Nutrition
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• v.25 no.4
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• pp.708-714
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• 1996

Ethanol production from raw starch was performed using the co-immobilized culture system of Rhizopu japonicus and zymomonas mobilis(R-Z). Glucose Production in immobilized R. japonicus culture was 2-fold higher than that in free cell culture. Ethanol production was 1.67g/L(Yp/s, 0.094) and 6.549/L(Yp/s, 0.38) in R-Z and R-Z 24 culture system, respectively. R-Z system was modified and designated as R-Z 24 system by replacing cotton plug with silicon check valve after 24h fermentation with R-Z system. Optimal substrate concentration for ethanol production in batch culture was 5%(w/v) and ethanol concentration produced was 15.02g/L(Yp/s, 0.36). Ethanol yield(Yp/s, 0.38) in fed-batch culture of 5 times with 2%(w/v) substrate was equal to that in batch culture of 2%(w/v) substrate.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.72-72
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• 2002

조류는 발생학적 특성상 수정과 초기 배발생을 제외한 거의 대부분의 개체발생 과정이 난각 속에서 진행된다. 그러므로 수정란에 생명 공학적 기법을 적용하는데 있어서, 포유류의 경우 여러 생명공학 기법이 적용된 수정란은 초기발생을 위한 체외배양 이후 반드시 모체에 이식되어야 하지만, 조류의 경우 인공적인 체외배양 체계가 확립되어 있어야 생명공학 기법이 적용된 수정란을 개체까지 발생시킬 수 있게 된다. 닭의 난자는 난관 누두부에 배란 후 약 15분내에 정자의 침입을 받아 수정되어 난관 팽대부에 도달하면 1세포기 수정란이 된다. 그 후 수정란이 협부에 도달하면 최초로 분할이 일어나기 시작하여, 방란시에는 그 세포수가 60,000개에 이르게 된다. 한편, 계란의 형성 과정에서는 다량의 난황을 포함한 난자가 난관 누두부로 배란되어 정자와 만나게 되면 수정란으로서 계란 형성이 계속되고 정자와 만나지 못하게 되면 무정란으로서 계란 형성을 계속하게된다. 배란된 난자가 난관누두부를 거쳐 난관팽대부에 도달되면 난자는 농후난백에 의해 둘러싸이게 되고 난관혈부에 도달되면 난각막이 형성되고 수양성 난백이 침적하게 된다. 그 후, 난관협부를 지난 난자는 난관자궁부에 도달되면 20시간이상 그곳에 머물면서 난각형성이 진행된 다음 방란된다. 따라서 수정란에 외래유전자를 미세주입하여 형질전환 닭을 생산하기 위해서는 수정란을 암탉의 난관 내에서 최초 분할되기 전에 외부로 끄집어내어야 하며, 수정란에 외래 유전자를 미세주입한 다음에는 다시 암탉의 난관내로 이식해야 하지만 현재까지 그러한 기술은 확립되어 있지 못하다. 그렇기 때문에 모체의 난관 속에서 일어나는 배 발생과 그 이후 개체까지의 발생을 위하여 인공적인 체외배양 체계가 확립되어 있어야 한다. 따라서 본 발표에서는 형질전환 닭을 생산하기 위한 양질의 1세포기 수정란 획득 방법과 획득된 수정란의 체외 배양방법에 관하여 기술적인 측면에서 고찰 해보고자 하며, 그와 같은 배양 기술을 이용하여 외래유전자를 도입한 일련의 결과에 관하여 보고 하고자한다.

• Journal of Embryo Transfer
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• v.14 no.2
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• pp.89-92
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• 1999

효율적인 homozygous 동물을 생산하기 위한 실험의 단계로 염색체가 4배체인 수정란의 이용성을 타진하기 위해 생쥐 수정란과 cytochalasin B를 사용하여 4배체 유도에 관한실험을 수행하였다. 생쥐 2-세포기 수정란을 10$\mu\textrm{g}$/ml 농도의 cytochasin B로 약 20시간 배양하였을 때 모든 수정란은 발육을 거의 멈추었으나, 이 수정란을 cytochalasin B-free medium에 체외배양하였을 때 발육이 재개되어 48시간 후 상실기나 배반포기까지 약 74%의 발육율을 나타내었다. 그러나 발육된 수정란의 세포수는 대조구에 비해 휠신 적은 것으로 나타났다. 염색체 분석결과 cytochalasin B로 처리한 대부분의 수정란은 4배체인 것으로 나타났고 약간의 수정란은 mosaicism과 다배체를 나타내기도 하였다. 그러므로 cytochalasin B를 이용하여 효과적으로 4배체의 수정란을 유도할 수 있는 것으로 나타났다.

• KSBB Journal
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• v.17 no.1
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• pp.110-113
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• 2002

Studies were made to examine the various effects of jasmonic acid and benzoic acid on ginsenoside production in suspension cultures of Panax ginseng C. A. Meyer. Jasmonic acid increased the ginsenoside production when it was dosed at the concentration of 50 $\mu$M or higher. The cell growth, however, was reduced with jasmonic acid. When benzoic acid was dosed simultaneously with iasmonic acid, the ginsenoside production increased 9.6 folds. It was 2.2 times higher than the result of single dose of jasmonic acid.

• Korean Journal of Plant Tissue Culture
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• v.25 no.1
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• pp.1-5
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• 1998

This study describes the effect of the growth regulators such as 2,L-D and BA, on the structural abnormalities of somatic embryos derived from leaf explants of Angelica gigas Nakai, Also, the relationship between the cotyledon number of a somatic embryo and its germinability is explored. Embryogenic calli were selected from calli formed on explants cultured on MS solid basal medium supplemented with 0.5mg/L 2,4-D, 1mg/L 2,4-D, 1mg/L plus 0.1mg/L BA, and 1 mg/L 2,4-D plus 0.5mg/L BA. Cotyledonary abnormalities were observed in somatic embryos which were developed from embryogenic calli cultured on MS medium containing 1mg/L 2,4-D for 8 weeks and then subcultured on 2,4-D free MS medium for 3 weeks. The frequency of abnormalities was as follows: 22.8% one cotyledon, 42.5% two cotyledons, 16.8% three cotyledons, 7.8% four cotyledons, 1.8% five cotyledons, and 8.2% jar shaped cotyledon. In addition, ABA treatment indicated an improvement of the somatic embryo with normal cotyledon (65.3%). ABA was important role to the high production of normal somatic embryos. Two cotyledon embryos showed germinability 77.8%. However the germinability of somatic embryos with anomalous cotyledons was prominently low: One cotyledon, 62.5%; three cotyledons, 43.3%; four cotyledons, 60%; five cotyledons, 50% and jar shaped cotyledon, zero%. Thus, germinability was essentially, inversely proportional to cotyledon number.

• Journal of Plant Biology
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• v.39 no.1
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• pp.71-77
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• 1996

Embryogenic callus was selected from callus induced from hypocotyl segment cultures of Daucus carota seedlings on MS basal medium supplemented with 1.0 mg/L 2,4-D. Cell clumps prepared from the embryogenic callus were transferred to MS medium without 2,4-D for somatic embryo development. Cotyledonary abnormalities were frequently observed on somatic embryos developed after the incubation of cell clumps in MS basal medium with 1.0 mg/L 2,4-D for one week and then subculture in the same medium but without 2.4-D for two weeks. The percentage of abnormalities was as follows: 5% one cotyledon, 21% three cotyledons, 6% four cotyledons, 5% five cotyledons, 0.2% six cotyledons and 1% trumpet-like cotyledons. On the other hand, the normal somatic embryo with two cotyledons appeared at 63%. The germination rate of somatic embryos was higher in two cotyledon somatic embryos than in multicotyledonary embryos. Trumpet-like somatic embryos did not germinate normally showing limited elongation and enlargement of roots and cotyledons without shoot development. From anatomical examination circular procambium in the root of somatic embryo began to branch around the middle regions of the hypocotyl which extended into the cotyledons through the cotyledonary nodes and the number of branched procambial strands in hypocotyl was equal to the number of cotyledons. Monocotyledonous somatic embryo always had larger cotyledon than that of somatic embryos with multicotyledons and had horseshoe-like cotyledons where the procambium was of the same structure.ucture.

• Korean Journal of Animal Reproduction
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• v.27 no.2
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• pp.125-133
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• 2003

The present study was carried out to examine the effect of $\beta$-Mercaptoethanol ($\beta$-ME) on in vitro maturation (IVM) of porcine follicular oocytes and oxygen concentration with $\beta$-ME on in vitro development (IVD) of porcine IVM/IVF embryos. The results were summarized as follows. 1. The rates of nuclear maturation, penetrated oocytes, polyspermic oocytes, pronucleus formation and mean numbers of the penetrated sperms were not significantly different using NCSU-23 maturation media for 0, 25, 50 and 100 $\mu$M $\beta$-ME (P>0.05). 2. The rates of blastocyst formation at day 7 after in vitro fertilization were higher in oocytes matured with 25 $\mu$M $\beta$-ME (25.4$\pm$0.9%) than in those matured with 0 (14.5$\pm$1.6%), 50 (17.3$\pm$1.7%) and 100 $\mu$M (12.4$\pm$1.3%) (P<0.05). However, no differences ware found in total cell numbers of blastocyst among the treatments. 3. The rates of blastocyst formation at day 7 after in vitro fertilization were higher in the NCSU-23 Culture medium With 25 $\mu$M $\beta$-ME (23.6$\pm$2.8%) than in those Cultured With 0 (15.4$\pm$4.4%), 12.5 (17.5$\pm$2.3%) and 50 $\mu$M $\beta$-ME (18.6$\pm$2.1%) Under the 5% and 20% $O_2$ Concentrations (P<0.05). However, no differences was found in total cell numbers of blastocyst among the treatments. These results suggested that the addition of 25 $\mu$M $\beta$-ME in the IVM/IVD media were effective on the porcine embryo production. However, the rates of blastocyst formation and total cell numbers of blastocyst at day 7 of porcine IVM/IVF embryos were not significantly different in the NCSU-23 culture medium under 5% and 20% 02 concentrations.