• Journal of Plant Biotechnology
• /
• v.35 no.1
• /
• pp.55-61
• /
• 2008

Previous studies on genetic transformation of chrysanthemum using cold regulated gene (BN115) have been conducted and the PCR and Real-Time PCR based method to determine the presence of the transferred cold regulated gene in the chrysanthemum was established. To check whether over-expression of BN115 gene in transgenic chrysanthemum will enhance their tolerance to cold stress, the transgenic chrysanthemum were grown under low temperature condition and several cold signalling including growth characteristics, stoma size and shape, SPAD value and ion leakage test were investigated. The transgenic chrysanthemum in the low temperature growth chamber grow much faster in term of the height, number and size of the leaves than those of wild-type plants and damage of transgenic plant caused by the low temperature was much less than that of wild-type plants. The stoma type and size of transgenic plant leaves grown at $5^{\circ}C$ were much similar to of wild-type plant cultured on $25^{\circ}C$ It has been found that SPAD value of transgenic plants was much higher than those of wild-type, but the EC density being lower under low temperature condition.

• Current Research on Agriculture and Life Sciences
• /
• v.5
• /
• pp.52-58
• /
• 1987

The yeast S. cerevisiae DBY747 was transformed with E. C - S. C shuttle vector YIp5, YEp13 and YRp7 by the method of spheroplast. The transformation frequency of YEp13 and YRp7 in S. cerevisiae DBY747 was $1.2{\times}10^3$ and $1.0{\times}10^2$ per $10{\mu}g$ of DNA, respectively. The transformants with YIp5 plasmid incapable of autonomous replication in S. cerevisiae were not detected in the condition of this experiment, but YIpS plasmid expressed the gene carried on it when cotransformed with a helper plasmid such as YEp13 or YRp7 : autonomously replicating plasmid. When plasmids were used in covalently closed circular form, cotransformation frequency of Ylp5-YEpl3 and Ylp5-YRp7 was 210 and 95 per $10{\mu}g$ of DNA, respectively. In cotransformation of linear plasmids, transformation frequency of the same cohesive ends was similar to that of noncomplementary cohesive ends. Transformants by the cotransformation with circular plasmids have been shown much higher frequency than with linear plasmids in S. cerevisiae DBY 747. The mitotic segregation stability test suggested that the cotransformant of YIpS-YEp13 was more stable than that of YIpS-YRp7.

• 한국생물공학회:학술대회논문집
• /
• 2003.04a
• /
• pp.284-287
• /
• 2003

Effects of ultrasound on cell growth and the production of human granulocyte-macrophage colony-stimulating factor (hGM-CSF) were investigated using transgenic Nicotiana tabacum cell suspension cultures. The culture suffered a slight growth depression immediately after the sonication, but gradually recovered to normal growth within 2 days. When the cells were exposed to ultrasound, the level of secreted hGM-CSF was 2.14 times higher than that in normal condition. From the beginning to 6 days of culture, production of secreted hGM-CSF was higher than that of control and then decreased. At the end of culture, however, hGM-CSF was considerably increased up to 36.7%. In the case of intracellular hGM-CSF, the level was slightly higher than that obtained in normal condition. Total hGM-CSF production was 31.5% higher than that of control culture after 6 days. The highest amount of hGM-CSF was $34.9\;{\mu}g/L$.

• 한국생물공학회:학술대회논문집
• /
• 2003.04a
• /
• pp.289-292
• /
• 2003

Effects of immobilization on the production of human granulocyte-macrophage colony-stimulating factor (hGM-CSF) by Nicotiana tabacum cells were investigated using alginate and polyurethane foam as immobilization matrices. Encapsulation of the cells in alginate decreased protein production by 50% compared with that of suspension culture. Maximum hGM-CSF concentration was obtained by the cells immobilizaed in polyurethane foam. High hGM-CSF production could be possible when polyurethane foam was used because of high specific production and easy immobilization for cell recycling process with high cell density.

• The Korean Journal of Zoology
• /
• v.35 no.3
• /
• pp.372-382
• /
• 1992

당뇨병유발제인 streptozotocin이 생쥐 신장 사구체곁세포의 미세구조에 어떠한 영향을 미치는 지를 알아보고자 일반계통인 ICR생쥐와 유전성 당뇨병계통인 KK생쥐에 streptozotocin을 투여하여 경시적으로 각 동물의 신장 사구체곁세포의 미세구조의 변화를 관찰하였다. Streptozotocin을 투여한 ICR생쥐의 사구체곁세포는 3일째부터 과립형질내세망의 미약한 팽창과 과립내에 대소 공포의 출현 및 용해소체가 간혹 관찰되었다. 그후 시간이 지남에 따라 더욱 심하여 특히 2주 및 4주에서는 과립형질내세망의 팽창, 사립체, 골지장치 및 리보소곤 등이 소수 출현하였는데 비해 대소 용해소체는 많이 관찰되었으며 심한 탈과립으로 인해 세포질내 과립의 면적이 현저히 감소되었다. 그러나 KK생쥐의 실험군에서는 전 실험군에 걸쳐 퇴행성변화가 적었으며 ICR 생쥐 실험군에 비해 그 영향이 훨씬 적었다. 이상의 결과를 종합하여 보면 정상 ICR생쥐에 streptozotocin을 투여하자 되면 ICR생쥐 사구체곁세포에서 과립의 유의한 감소 및 세포내 미세구조의 퇴행성변화가 뚜렷한데 비해 KK생쥐 실험군에서는 ICR생쥐 실험군에 비해 손상을 적게 받았는데 이는 KK생쥐가 갖고 있는 당뇨병에 대한 내성에 의해 영향을 적게 미치는 것이 아닌가 추측된다.

• Proceedings of the Korea Society of Poultry Science Conference
• /
• 2004.11a
• /
• pp.9-10
• /
• 2004

We developed a new panel of markers for the characterization of chicken PGCs. The results of immunostaining demonstrated that anti-SSEA-3, anti-SSEA-4, anti-integrin 6, and anti-integrin 1 antibodies. and STA and DBA bound specifically to chicken PGCs. These reagents could be used to characterize chicken PGCs together with conventional marker reagents such as PAS and anti-SSEA-1 antibody. We also showed that double staining of PGCs with the newly developed markers was feasible, which might contribute to rapid detection and accurate characterization of chicken PGCs.

• Proceedings of the KSAR Conference
• /
• 2001.03a
• /
• pp.57-57
• /
• 2001

Erythropoietin(EPO)는 혈액의 구성성분 중에서 적혈구 세포 증식에 중요한 기능을 한다고 알려져 있으며, 최근에는 암, 에이즈의 치료 등에도 효과가 있는 것으로 확인되고 있다. 따라서 본 연구팀은 지금까지 hEPO 유전자를 이용하여, 형질전환 돼지 "새롬이"의 생산에 성공한바 있다. 새롬이의 정액을 활용하여 인공수정을 실시 새롬이의 Fl를 24두 생산하였다. 이에 대하여 "형질전환 돼지의 계대번식시 유전자 전이효율에 관한 연구"라는 제목으로 발표 할 예정이며, 형질전환에 사용된 promoter가 WAP이므로, Fl이 임신, 분만을 하여야만 유즙을 통하여 hEPO물질을 생산할 수 있다. 따라서, Fl(♂)$\times$Fl(♀)의 교배에 의하여 5두가 임신, 분만을 하였으며, 이들 중 1두는 분만 후 21일에 폐사하였으며, 나머지는 현재 정상적으로 사육되고 있다. 이들 5두에 대하여 분만 후 유즙을 채취하여 유즙속에 EPO의 발현여부를 검토하였다. EPO-ELISA kit(medac)를 사용하여 분석결과, 유즙을 8,000배로 희석을 하여야만 Standard curve(1.25~160 mIU/$m\ell$)안에서 EPO의 단백질 발현을 검출할 수 있었다. 5두의 각각 농도는 28, 58, 17, 37, 27 IU/${\mu}\ell$ 였다. 또한 cDNA EPO와 genome EPO를 CHO 동물세포에서 생산하여 10배로 농축한 결과 5.5와 11 IU/${\mu}\ell$의 농도로 유즙과 비교하면 약 20~30배의 낮은 발현양을 나타내었으며, 또한 이러한 결과는 소변에서의 결과(1.1 IU/$m\ell$)보다는 약 30,000배 이상 높은 발현량을 화인 할 수 있었다. 현재, 이들 유즙 물질을 활용 빈혈 질환실험동물을 이용하여 생리활성을 검정, 체내에서 metabolic clearance rate(MCR)를 검토 중에 있다. 또한 F2의 자돈생산은 모돈 5두에서 총 25두가 생산되었는데, 이중 20두 약 80%가 EPO 유전자의 전환율을 나타내었다. 이상을 종합하면, 1) 돼지이용 생리활성물질(EPO)을 유즙에서 대량으로 생산할 수 있는 system의 활용가능성을 국내에서 처음으로 확인하였으며, 2) EPO에 있어서는 국제적으로도 형질전환 가축생산은 최초로 성공하였으며, 현재로서는 생산되어진 물질의 정제수준에 따라 활용가치가 결정되어 질 것으로 사료된다. 생리활성 물질을 생산할 수 있는 형질전환 돼지 생산의 성공은, 앞으로 형질전환 가축생산 뿐 만 아니라, 장기이식 및 복제돼지 생산의 활용 면에서의 응용가능성이 기대된다.

• Journal of Plant Biotechnology
• /
• v.33 no.3
• /
• pp.185-194
• /
• 2006

Chloroplasts are believed to be descended from certain cyanobacteria, which were taken up by phagocytosis into a host cell and lived there in a symbiotic relationship. In contrast to the current static concept on the chloroplast genome, its dynamism has been recently demonstrated: the chloroplast genome is active in intramolecular homolgous recombination, producing subgenomic circles when it obtains homolgous sequences via genetic transformation. Chloroplast tranformation in higher plants provides many advantages over nuclear transformation that include higher expression levels of transgenes, polycistronic expression of transgenes, and maternal transmission of transgenes. Tobacco has been used as a model for chloroplast genetic transformation. However, it is recently possible to transform the chloroplasts of other major food and economic crops including rice, soybean, and cotton. Chloroplast-transformed crops will be able to replace bioreactors using microorganisms for production of value-added proteins in future.

• Korean Journal of Plant Tissue Culture
• /
• v.22 no.4
• /
• pp.235-240
• /
• 1995

The development of selectable markers for transformation has been a major factor in the successful genetic manipulation of plant. We established a new selectable marker system for tobacco transformation using chimeric adenosine deaminase (ADA) gene, which confers resistance to cytotoxic adenosine analogues, 9-$\beta$-D-arabinofuranosyl adenine(Ara-A) and cordycepin. The transformants with the chimeric ADA gene in tobacco grew in the presence of normally lethal level of cytotoxic adenosine analogues, 100 $\mu$M Ara-A and 50 $\mu$M cordycepin. We successfully distinguished transformed shoot from non-transformed shoot on the same selectable media with cytotoxic adenosine analogues. In this selectable media, we were able to select seeds with/ without ADA gene from transgenic tobacco seeds. Theses results show that the mammalian ADA gene may serve as a new selectable marker for tobacco transformation.

• Journal of the Korean Society of Radiology
• /
• v.83 no.4
• /
• pp.924-930
• /
• 2022

Most spinal meningiomas have an intradural or partly extradural location. The meningothelial origin is the most common pathologic type of spinal meningioma. Pure extradural spinal meningiomas are not common, and lymphoplasmacyte-rich meningioma (LPRM) is very rare. We report a case of isolated extradural spinal meningioma in the thoracic spine that was pathologically confirmed as LPRM.