• 한국수산과학회지
• /
• 제32권1호
• /
• pp.117-120
• /
• 1999

해산 녹조류 참흩파래, Monostroma nitidum의 엽체를 효소처리하여 다량의 원형질체를 분리하였다. 최적 효소액의 조합은 $4\%$ R-10+$3\%$ Macerozyme R-10+$3\%$ Abalone acetone power로서 생체조직 300mg 당 $4.41\times10^6$개의 원형질체가 분리되었다. 원형질체의 수율은 효소처리 270분에 최대였다. 분리 직후의 원형질체는 구형으로 직경 $13\~33\mu$m의 크기였다. 분리된 원형질체는 0.4M mannitol을 함유한 f/2배지에서 배양한 후 매주 mannitol이 함유되지 않은 f/2 배지로 절반씩 교환함으로서 분화율을 높일 수 있었다. f/2배지를 사용한 적정 배양조건에서 원형질체는 배양 3일 후 새로운 세포벽을 형성하였으며 10일 후 발아하기 시작하여 엽체로 발달하였다. 항생물질의 배지내 첨가는 배양체의 분화를 저해하였다.

• 한국수정란이식학회지
• /
• 제11권1호
• /
• pp.15-26
• /
• 1996

This study was conducted to improve the in vitro maturation(JVM), in vitro fertilization (IVF) and in vitro developmental capacity of oocytes derived from slaughtered Korean native cattle. The recoverd oocytes, obtained from a local slaughter house, were used completely surrounded by at least 3 layers of cumulus cells in combination with a homogeneous cytoplasmic pigmentation. In vitro maturation was induced in TCM-199 or Ham's F-10 supplemented with LH(1O $\mu$g/rnl), FSH(35 $\mu$g/ml), estradiol-17$\beta$(1 $\mu$g/ml) at 39$^{\circ}C$ under 5% $CO_2$ in air for 24 hours. Sperm from caudal epididyrnis and previously matured cumulus-oocytes complexes were cultured for 24 hours in 100 $\mu$l droplets of fertilization media under paraffin oil. The zygotes were cultured with media(TCM-199 with bovine oviductal epithelial cells or CRlaa) for 7 to 10 days. The cleavage rate of IVM-IVF oocytes was significantly (P<0.05) higher following maturation using Ham's F-10 (59.9%) than TCM-199 (51.6%). Development to the blastocysts among cleaved embryos was not signficantly different between maturation media: Ham's F-10 (16.0%) and TCM-199(11.9%). However, the hatching rate was affected significantly (P<0.05) on rnaturation media as 62.9% in Ham's F-10, compared with 41.2% in TCM-199. The cleavage rate of IVM-IVF oocytes was significantly (P<0.05) higher following IVF using m-TALP medium (80.1%) than BO medium (51.6%). The percentage of in vitro developed blastocysts among cleaved embryos was not signficantly different between fertimization media: BO (11.7%) and m-TALP (17.6%). The cleavage and the developmental rate to the blastocysts after IVF in m-TALP or condition medium(CM) with or without oviduct epithelial cell monolayer(OECM) was similar(80.1% and 17.6% in m-TALP, 83.8% and 19.4% in M-TALP with OECM. 82.9% and 18.9% in CM, 87.6% and 16.0% in CM with OECM, respectively). The percentage of in vitro developed blastocysts among cleaved embryos was significantly (P<0.05) higher in TCM-199 medium co-cul tured with bovine oviduatal epithelial cell monolayers(35.2%) than CRlaa medium(1.9%). These results stggest that the most transferable IVF embryos could be produced from Ham's F-10, m-TALP and TCM-199 medium with bovine oviductal epithelial cell monolayers for IVM, IVF and IVC, respectively.

• Journal of Microbiology and Biotechnology
• /
• 제10권6호
• /
• pp.789-796
• /
• 2000

In order to analyze the effects of tktA, $aroF^{FBR}$, and aroL expression in a tryptophan-producing Escherichia coli, a series of plasmids carrying the genes were constructed. Introduction of tktA, $aroF^{FBR}$, and aroL into the E. coli strain resulted in approximately 10-20 fold increase in the activities of transketolase, the feedback inhibition-resistant 3-deoxy-D-arabinoheptulsonate-7-phosphate synthase, and shikimate kinase. Expression of $aroF^{FBR}$ in the aroB mutant strain of E. coli resulted in the accumulation of 10 mM of 3-deoxy-D-arabinoheptulsonate-7-phosphate (DAHP) in the medium. Simultaneous expression of tktA and $aroF^{FBR}$ in the strain further increased the amount of excreted DAHP to 20 mM. In contrast, the mutant strain which has no gene introduced accumulated 0.5 mM of DAHP. However, the expression of tktA and $aroF^{FBR}$ in a tryptophan-producing E. coli strain did not lead to the increased production of tryptophan, but instead, a significant amount of shikimate, which is an intermediate in the tryptophan biosynthetic pathway, was excreted to the growth medium. Despite the fact that additional expression of shikimate kinase in the strain could possibly remove 90% of excreted shikimate to 0.1 mM, the amount of tryptophan produced was still unchanged. Removing shikimate using a cloned aroL gene caused the excretion of glutamate, which suggests disturbed central carbon metabolism. However, when cultivated in a complex medium, the strain expressing tktA, $aroF^{FBR}$, and aroL produced more tryptophan than the parental strain. These data indicate that additional rate-limiting steps are present in the tryptophan biosynthetic pathway, and the carbon flow to the terminal pathway is strictly regulated. Expressing tktA in E. coli cells appeared to impose a great metabolic burden to the cells as evidenced by retarded cell growth in the defined medium. Recombinant E. coli strains harboring plasmids which carry the tktA gene showed a tendency to segregate their plasmids almost completely within 24h.

• Journal of Ginseng Research
• /
• 제31권4호
• /
• pp.210-216
• /
• 2007

The backbone structure of ginsenosides, active ingredients of Panax ginseng, is similar with that of sterol, especially cholesterol. Caenorhabditis elegans (c. elegans) is one of free living nematodes and is well-established animal model for biochemical and genetic studies. C. elegans cannot synthesize de novo cholesterol, although cholesterol is essential requirement for its growth and development. In the present study, we investigated the effects of Korean red ginseng total extract (KRGE), ginseng total saponins (GTS) on life span of C. elegans in cholesterol-deprived and -fed medium. Cholesterol deprivation caused damages on life span of worms throughout F1 to F3 generations. KRGE or GTS supplement to cholesterol-deprived medium restored the life span of worms as much as cholesterol alone-fed medium. In study to identify which ginsenosides are responsible for life span restoring effects of KRGE, we found that ginsenoside Rc supplement not only restored life span of worms grown in cholesterol-deprived medium but also prolonged life span of worms grown in cholesterol-fed medium. These results show a possibility that ginsenosides could be utilized by C. elegans as a sterol substitute and further indicate that ginsenoside Rc is the effective component of Korean red ginseng that prolongs the life span of C. elegans.

• KSBB Journal
• /
• 제22권5호
• /
• pp.313-317
• /
• 2007

본 연구에서는 Pluronic F-68을 해동 후 회복배지에 적용하여, 동결보존된 형질전환 식물세포의 생장 증진을 도모하였다. Pluronic F-68은 세포표면의 소수성을 감소시켰을 뿐만 아니라, 세포의 투과성을 증진시켜 세포와 직접 상호 작용할 수 있음을 관찰하였다. 또한 Pluronic F-68의 첨가에 의해 재조합단백질의 발현에 부정적인 영향을 보이지 않음을 확인하였다. 따라서 동결보존시 해동 후 회복배지에 Pluronic F-68의 첨가는 식물세포의 신속하고 효율적인 대량 현탁배양을 가능하게 할 수 있다.

• 미생물학회지
• /
• 제28권3호
• /
• pp.274-277
• /
• 1990

새로운 고체배지 응고제인 Pluronic Polyol F127을 한천대신 사용하여 C. congregatus를 배양하였고 균사 끝(hyphaltip)에 있는 phenoloxidase들의 세포내에서의 존재위치를 확인하였다. 고체배지의 균사에서 원형질체를 얻었고 concanavalin A를 처리하여 세포막을 분리하였다. 세포막 분획에 phenoloxidase들이 존재하였고 이는 이 효소가 light receptor complex의 한 부분임을 시사한다.

• Fisheries and Aquatic Sciences
• /
• 제14권4호
• /
• pp.317-322
• /
• 2011

This study was conducted to develop economical agricultural fertilizer media for the mass culturing of Nannochloropsis oceanica. Specific growth rates of N. oceanica cultured with differing concentrations of commercial compounds, urea fertilizers, and trace elements (Zn, Cu, Co, Mo) were compared with the growth rate in f/2 medium. Among the various added trace elements, $CuSO_4{\cdot}5H_2O$ was most effective for high growth of N. oceanica. The main nitrogen source in the agricultural fertilizers was ammonium, which was unsuitable for the growth of N. oceanica. Thus, the fertilizer at a lower concentration infused with $NaNO_3$ as a nitrogen source was more effective than fertilizer at higher concentrations. In this study, the growth of N. oceanica cultured with an agricultural fertilizer medium composed of compound fertilizer (41.7 mg/L), urea fertilizer (34.4 mg/L), $NaNO_3$ (150 mg/L), and $CuSO_4{\cdot}5H_2O$ (0.0588 mg/L) was similar to that of N. oceanica cultured in f/2 medium.

• 한국수정란이식학회지
• /
• 제15권1호
• /
• pp.39-46
• /
• 2000

This study was conducted to investigate the effect of hormones, protein sources and anti-oxidants on in vitro maturation (IVM) and in vitro fertilization(IVF) of bovine follicular oocytes. The rates of Holstein follicular oocytes classified as grade A and B(50.2% and 33.2%) were higher than those of Hanwoo cattle(40.3% and 32.0%, P<0.05). The cumulus cell expansion rates of oocytes cultured in TCM-199 and Ham's F-10 medium supplemented with 10% FCS and hormones were higher (81.9~87.6%) than those of non-treated groups (74.5~81.7%). The fertilization rates of oocytes cultured in TCM-199 and Ham's F-10 medim supplemented with 10% FCS, 1% BSA and 10% bFF was 53.8~55.0%, 51.4~52.6%, and 47.0~50.0%, respectively. The polyspermy rates was 13.6~14.2%, 10.0~11.1%, and 10.0%, respectively. When the oocytes were cultured in TCM-199 and Ham's F-10 medium with 50${\mu}{\textrm}{m}$ $\alpha$-tocopherol, the fertilization rates was 62.0 and 60.2%, respectively. In the maturation medium added of 100${\mu}{\textrm}{m}$ cysteamine, the fertilization rates was 64.7 and 66.7%, respectively. The fertilization and polyspermy rates of treated groups were higher than those of non-treated group. The results show that hormones, protein sources and anti-oxidants can provide a benefit for in vitro maturation and fertilization of bovine follicular oocytes.

• 한국환경과학회지
• /
• 제19권9호
• /
• pp.1077-1082
• /
• 2010

This study was performed to investigate the nutritional conditions controlling keratinase activity in Bacillus megaterium F7-1. B. megaterium F7-1 produced keratinase using chicken feather as a sole source of carbon, nitrogen and sulfur. Addition of the feather medium with glucose enhanced keratinase production (68.9 U/ml), compared to control without glucose (63.2 U/ml). The synthesis of keratinase was repressed by addition of $NH_4Cl$ in B. megaterium F7-1. The highest keratinase production (70.9 U/ml) was obtained with the feather medium containing glucose and $MgSO_4{\cdot}7H_2O$. Keratinase was produced in the absence of feather (4.9 U/ml), indicating its constitutive synthesis. Feather degradation resulted in free SH group formation. B. megaterium F7-1 effectively degraded chicken feather meal (86%), whereas duck feather, human nail, human hair and sheep wool displayed relatively low degradation rates (8-34%).

• 대한한의학회지
• /
• 제24권2호
• /
• pp.159-165
• /
• 2003

Objectives : Coptidis rhizoma (CR) is an oriental medicine that has been used in many traditional prescriptions against diabetes mellitus in Korea for centuries. Our purpose was to determine the protective effect and its action mechanism of CR on the cytotoxicity of pancreatic -cell line (RINm5F cell). Methods : In this experiment, we used methods such as MTT assay for detection of cytotoxicity, DNA fragmentation assay for detection of apoptotic cell death, LDH activity assay for detection of necrotic cell death, and measurement of $DiOC_{6}$ (3) retention for detection of mitochondrial membrane potential (MMP). Background : Nitric oxide (NO) is believed to playa key role in the process of pancreatic -cell destruction leading to insulin-dependent diabetes mellitus (IDDM). Results : Exposure of RINm5F cells to chemical NO donor such as S-nitroso-N-acetylpenicillamine (SNAP) induced cytotoxic events such as DNA fragmentation and lactate dehydrogenase (LDH) release into medium. However, pretreatment of RINm5F cells with CR extract ($10~50{\mu\textrm{g}}/ml$) for 3 hours prevented SNAP-induced DNA fragmentation and LDH release into medium through the inhibition of MMP disruption. Conclusions : These results suggest that CR may be a candidate for a therapeutic or preventing agent against IDDM.