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배추의 형질전환용 선발항생제로서 Paromomycin의 이용
조미애,민성란,고석민,유장렬,이준행,최필선,Cho, Mi-Ae,Min, Sung-Ran,Ko, Suck-Min,Liu, Jang-Ryol,Lee, Jun-Haeng,Choi, Pil-Son 한국식물생명공학회 2006 식물생명공학회지 Vol.33 No.4
정상' 배추와 '서울' 배추의 배축절편을 선발마커로서 hygromycin 저항성유전자를 갖고 있는 pCAMBIA1301와 paromomycin저항성유전자를 갖고 있는 pPTN290으로 각각 형질 전환된 LBA4404 또는 EHA101균주와 공동배양한 후 선발배지에서 배양하면서 형질전환체를 선발하였다. 형질전환빈도는 사용된 항생제와 품종에 따라서 현저하게 차이가 있었으며, 특히 paromomycin은 hygromycin보다 효과적이었고 정상 배추는 서울배추보다 양호하였다. 가장 높은 형질전환빈도는 (0.70%) 100mg/L paromomycin이 첨가된 선발배지에서 정상배추의배축을 배양할 경우 얻어졌다. GUS양성반응으로 확인 한 결과 정상배추에서 9개체와 서울배추에서 3개체를 각각 얻었으며, 온실에서 생장한 후 $T_1$종자를 수확하였다. $T_1$ 종자를 다시 발아시켜 유식물체를 얻은 후 GUS양성반응을 확인함으로서 외래유전자가 안정적으로 발현하고 있음을 확인하였다. Hypocotyl explants of Chinese cabbage (us. 'Jeong Sang' and 'Seoul') produced adventitious shoots on Murashige and Skoog (MS) basal medium supplemented with 4mg/L $AgNO_3$, 5 mg/L acetosyringone, 4 mg/L 6-benzyladenine and 3mg/L alpha-naphthaleneacetic acid (SI) after cocoultivation with strains of Agrobacterium tumefaciens (LBA4404) harboring the pCAMBIA1301 and the $_PPTN290$ containing hygromycin-resistance gene and paromomycin-resistance gene as a selectable marker genes, respectively. There was a significant difference in the frequency of transgenic plants depending on antibiotics and cultivars used. Paromomycin was better than hygromycin, and cultivar 'Jeong-sang' was higher than 'c.v. Seoul' in the frequency of transgenic plants. In particular, the highest frequency (0.70%) of transgenic plants was obtained from selection medium (SI) containing 100mg/L paromomycin in c.v., 'Jeong-sang' GUS positive response were obtained 9 plants and 3 plants from the cultivars, 'Jeong-sang' and 'Seoul', respectively. They were grown to maturity in a greenhouse and normally produced $T_1$ seeds. GUS histochemical assay for progeny $(T_1)$ revealed that the transgenes were expressed in the plant genome.
배추의 배축절편으로부터 캘러스와 뿌리 발생을 통한 안정적 형질전환
조미애,김춘해,민성란,고석민,유장렬,최필선,Cho, Mi-Ae,Kim, Choon-Ae,Min, Sung-Ran,Ko, Suck-Min,Liu, Jang-Ryol,Choi, Pil-Son 한국식물생명공학회 2007 식물생명공학회지 Vol.34 No.2
'정상' 배추의 배축절편을 선발마커로서 paromomycin 저항 성유전자를 갖고 있는 pPTN290으로 각각 형질전환된 EHA101, LBA4404, GV3101균주와 공동배양한 후 갤러스유도배지에서 형질전환캘러스를 얻은 후, 뿌리유도배지에서 부정근을 그리고 신초유도배지에서 신초를 각각 순차적으로 유도하였다. 형질전환캘러스를 얻은 후, 뿌리유도배지에서 부정근을 그리고 신초유도배지에서 신초를 가각 순차적으로 유도하였다. 형질전환캘러스 형성은 Agrobacterium균주에 따라 차이가 있었으며, 특히 EHA101균주에 공동배양된 배축절편으로부터 최대 6.1%까지 얻어졌다. 또한 각각의 형질전환캘러스 클론으로부터 형질전환 부정근과 신초 발생은 EHA101균주에서 60.7%와 38.2%, LBA4404에서 8.3%와 0%, GV3101에서 20.5%와 85.7%까지 각각 얻을 수 있었다. 형질전환식물체는 특별한 형태적 이상 없이 온실에서 정상적으로 자라 $T_{2}$종자를 얻을 수 있었다. GUS방법으로 7개의 후대 유식물체를 분석한 결과 gus유전자가 안정적으로 발현하고 있음을 확인하였고, 배추 genome에 single 또는 multiple copy로 전달되고 있음을 추측할 수 있었다. Hypocotyl explants of Chinese cabbage (cvs. "Jeong Sang") produced transgenic calli on callus induction medium (MS salt, B5 vitamin, 5 mg/L acetosyringone, 1 mg/L 2,4-D, 3% sucrose, 400 mg/L cefotaxime, 100 mg/L paromomycin, pH 5.8) after cocultivation with strains of Agrobacterium tumefaciens (EHA101, LBA4404, GV3101) harboring the pPTN290 containing paromomycin-resistance gene as a selectable marker, and then they transferred to root induction medium (1/2MS salt, MS vitamins, 2% sucrose, 100 mg/L paromomycin, 100 mg/L cefotaxime, pH 5.8) and shoot induction medium (MS salt, B5 vitamin, 4 mg/L $AgNO_3$, 4 mg/L 6-benzyladenine, 3 mg/L alpha-naphthaleneacetic acid, 100 mg/L paromomycin, 100 mg/L cefotaxime, 3% sucrose, pH 5.8) in order. There was a significant difference in the frequency of transgenic calli depending on Agrobacterium strains. In particular, the highest frequency (6.1%) of transgenic calli was obtained from the hypocotyls cocultivated with EHA101 strains. Also, the frequency (%) of transgenic root and plants from each transgenic callus clone were obtained with 60.7% and 38.2% in EHA101, with 8.3% and 0% in LBA4404, with 20.5% and 85.7% in GV3101 strains, respectively. They were grown to maturity in a greenhouse and normally produced $T_2$ seeds. GUS histochemical assay for progeny ($T_2$) revealed that the transgenes was expressed in the plant genome, and progeny analysis from 7 independent transgenic events demonstrated that the transformants transmitted the transgene as a single or multiple functional locus.
( Sung Pil Choo ),( Min Jung Lee ),( Inha Lee ),( Jae Hoon Lee ),( Heeyon Kim ),( Gee Soo Jung ),( Sehee Kim ),( Wooseok Im ),( Young Sik Choi ),( Sihyun Cho ) 대한산부인과학회 2022 대한산부인과학회 학술대회 Vol.108 No.-
Objective: Particulate matter (PM) has broad toxic effects and numerous studies have suggested possible links between the recent decline in female fertility and increased PM exposure. However, the effects of PM exposures on female reproductive function are not fully understood yet. This study aimed to evaluate adverse effects of PM on endometrium and identify possible underlying mechanisms. Methods: Endometrial cells from the patients who underwent surgeries for benign conditions were obtained, cultured, and treated with 50~200 ug/ml of PM for 4, 8, and 12 hours, respectively. In vivo model, female C57BL/6 mice were exposed to urban particulate matter (low and high dose) via intranasal inhalation for four weeks. Primary cultured endometrial cells and mouse endometrium were harvested, and the expression levels of pro-inflammatory cytokines involved in mitogen-activated protein kinase (MAPK) signaling and apoptotic pathways were evaluated using qRT-PCR and western blot. Results: The expression levels of Bax/Bcl-2 ratio, Cas-3, and p53 were significantly higher in primary cultured human normal endometrial cells upon 12 hours of PM exposure. Furthermore, the mRNA levels of IL-1 were significantly upregulated after PM exposure (30.68 folds). A similar trend was observed in the endometrium of PM-exposed mice. A marked increase in the mRNA levels of IL-1 was observed in low dose(7.41 folds) and high dose (26.98 folds) PM exposed mice compared with that in the control group. Subsequently, we found that NF-kB activation leads to pro-inflammatory cytokine upregulation. In addition, Massons Trichrome staining showed increased intensity of collagen fiber content in PM exposed endometrium in a dose-dependent manner, which implies increased fibrosis of endometrium. Conclusion: In this study, we identified that exposure to PM induced adverse effects on endometrium via alteration of apoptotic and inflammatory pathways. Further, PM exposure may be associated with endometrium fibrogenesis (fibrosis).
Sediment Toxicity of Industrialized Coastal Areas of Korea Using Bioluminescent Marine Bacteria
Choi, Min-Kyu,Kim, Seong-Gil,Yoon, Sang-Pil,Jung, Rae-Hong,Moon, Hyo-Bang,Yu, Jun,Choi, Hee-Gu The Korean Society of Fisheries and Aquatic Scienc 2010 Fisheries and Aquatic Sciences Vol.13 No.3
The quality of marine sediments from the industrialized coastal areas of Korea (Ulsan Bay, Masan Bay, and artificial Lake Shihwa) was investigated using a bacterial bioluminescence toxicity test. Sediment toxicity results were compared with the levels of chemical contamination (trace metals, organic wastewater markers, acid volatile sulfides, total organic carbon). Effective concentration 50% (EC50) of sediments ranged from 0.014 to 1.126 mg/mL, which is comparable to or lower than values in contaminated lakes, rivers, and marine sediments of other countries. Sediment reference index (SRI) ranged from 13 to 1044, based on the EC50 of the negative control sample. Mean average SRI values in Masan Bay and Lake Shihwa were approximately 8 and 9 times as high as that in Ulsan Bay, indicating higher sediment toxicity and greater contamination in the two former regions. Sediment toxicity were strongly associated with the concentrations of some chemicals, suggesting that this test may be useful for determining potential chemical contamination in sediments.