• Korean Journal of Plant Tissue Culture
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• v.24 no.2
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• pp.119-124
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• 1997

Plantlets were regenerated from various explants (shoot tip, leaf blade, petiole and root segments) via organogenesis and/or somatic embryogenesis from Armoracia rusticana(Lam) Gaerth., Mey, et Scherb.. Shoot regeneration rate from callus was highest on the MS mediums supplemented with 0.5 ㎎/L IAA, 5.0㎎/L BA and 10.0㎎/L spermine. A Low frequency of regeneration occurred on hormone-free MS medium. Multiple shooks were regenerated at a pH of 4.0 to 8.0 on MS medium supplemented with 1.0 ㎎/L BA and 0.1 ㎎/L NAA. Polyamines promoted shoot- and root-formation by 2 to 4 times normal, Specific proteins associated with organogenesis were identified. Somatic embryogenesis occurred directly from the leaf blade, petiole and root segments cultured on MS medium with 2.0 ㎎/L BA and 2.0 ㎎/L BA and 2.0 ㎎/L NAA. Three types of regeneration in A, rusticana were clearly established, which could be applied to the study of morphogenesis and genetics at cell, tissue and organ levels.

• Journal of Plant Biotechnology
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• v.42 no.4
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• pp.356-363
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• 2015

In order to study genetic engineering in trees, the characterization of genes and promoters from trees is necessary. We isolated the promoter region (867 bp) of Pagns-LTP from poplar (P. alba ${\times}$ P. glandulosa) and characterized its activity in transgenic poplar plants using a ${\beta}$-glucuronidase (GUS) reporter gene. High-level expression of the Pagns-LTP transcript was found in poplar roots, while comparatively low-level expression was found in the young leaves. Pagns-LTP mRNA was not detected in other poplar tissues. Additionally, transgenic poplar plants that contained a Pagns-LTP promoter fused to a GUS reporter gene, displayed tissue-specific GUS enzyme activity localized in root tissue. In silico analysis of the Pagns-LTP promoter sequence reveals the presence of several cis-regulatory elements responsive to phytohormones, biotic and abiotic stresses, as well as those regulating tissue-specific expression. These results demonstrate that the Pagns-LTP promoter has tissue-specific expression activity in poplar roots and leaves that may be involved in organ development and plant resistance to various stresses. Therefore, we anticipate that the Pagns-LTP promoter would be a useful tool to genetically optimize woody plants for functional genomics.

• Journal of Plant Biotechnology
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• v.34 no.3
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• pp.223-227
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• 2007

Plant regeneration system from leaf and root segments of Lycoris chejuensis via bulblet formation was established. Surface-sterilized leaf and root segments were cultured on the B5 medium containing 2,4-D. After 12 weeks of culture onto B5 medium containing 2,4-D, white globular structures and white calluses were formed on the cut surface of the explants. The highest frequency of globular structures and calluses formation from leaf explants was 32.1% when leaf explants were cultured onto B5 medium supplemented with 1 mg/L of 2,4-D. However, the higher concentration of 2,4-D (over than 3 mg/L) resulted in decrease of the frequency. In comparison to leaf explants, root segments showed the highest frequency at a rate of 36.1% when root explants were cultured onto B5 medium supplemented with 3 mg/L of 2,4-D. These structures and calluses were sub-cultured and proliferated onto the same culture medium. Upon transfer to B5 basal medium, white globular structures were developed into bulblets and normal plantlets. After 4 weeks of incubation in the light, plantlets were successfully rooted over the frequency of approximately 90%. Rooted plantlets were successfully transferred to potting soil and acclimatized in the growth chamber. The plant regeneration system of Lycoris chejuensis established in this study, might be applied to mass proliferation, conservation of genetic resources and genetic transformation for molecular breeding.

• Journal of Sericultural and Entomological Science
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• v.52 no.1
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• pp.39-44
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• 2014

We isolated highly-expressed genes in the posterior silk glands of silkworm on a previously study, which one of these was identified as RNA binding protein-1 homologue (RBP-1) gene. In this study, we investigated gene expressional characteristics of the RBP-1 depending on silkworm development stages and several tissues of the larvae, respectively. Northern blot hybridization analysis showed that the RBP-1 gene was expressed high in larval and pupal periods, and highly expressed than endogenous internal control gene (BmA3) on all tested larval tissues. In addition, we isolated and analyzed a phage DNA having 1,660 bp-long promoter region of the RBP-1 gene from a genomic DNA library. To study the RBP-1 gene promoter activity, RBP-1 (-740/+ 30) was amplified by PCR and subcloned into a pGL3 basic vector to generate pGL-RBP1. A luciferase report vector carrying RBP-1 gene promoter (770 bp) was tested by luciferase assay in Sf9 cells. In the result, the RBP-1 gene promoter was more efficient than constitutive promoter (BmA3) by approximately ten percent.

• Microbiology and Biotechnology Letters
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• v.33 no.4
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• pp.267-273
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• 2005

To develop effective and powerful probiotic, Saccharomyces cerevisiae strains producing cellulolytic enzymes were genetically brooded. For the production of exoglucanase, the plasmid pVT-TExo (8.8 kb) was constructed, in which Thermomonosporafusca exoglucanase gene (E3) was under the control of ADHl promoter, and introduced into S. cerevisiae SEY2102. When the transformant, S. cerevisiae SEY2102/pVT-TExo, was cultivated on YPD medium, the total expression level of avicelase reached about 190 unit/l. The secretion efficiency and plasmid stability were about $50\%\;and\;91\%$, respectively. Recombination exoglucanase enzyme bound to avicel better than Clostridium endoglucanase (CelA) and Trichoderma endoglucanase (C4) enzymes. The mixing ratio of E3 and CelA displaying the best synergistic hydrolysis for avicel was observed at 4:1. The mixture of endoglucanase (CelA) and exoglucanase (E3) resulted in 3.2-fold increase of avicelase activity and 2.5-fold enhanced production of sugar production from avicel, compared to the single enzyme treatment.

• Microbiology and Biotechnology Letters
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• v.39 no.4
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• pp.337-344
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• 2011

Integrase MJ1 from the bacteriophage ${\Phi}FC1$ carries out recombination between two DNA sequences (the phage attachment site, attP and the bacterial attachment site, attB) in NIH3T3 mouse cells. In this study, the integration vector containing attP, attB and the integrase gene MJ, was constructed. The integration mediated by integrase MJ1 in Escherichia coli led to excision of LacZ. Therefore, the frequency of integration was measured by the counting of the white colony, which is detectable on X-Gal plates. The extrachromosomal integration in NIH3T3 mouse cells was monitored by the expression of the green fluorescent protein (GFP) as a reporter. To demonstrate integration mediated integrase MJ1 in NIH3T3 cells, vectors containing attP and attB were co-transfected into NIH3T3 cells. The integration was confirmed by fluorescent microscopy. The expression of GFP was induced in NIH3T3 cells expressing MJ1 without accessory factors. By contrast, the excision mediated by the MJ1 between attR and attL had no effect on the expression of GFP. These results suggest that integrase MJ1 may enable a variety of genomic modifications for research and therapeutic purposes in higher living cells.

• Korean Journal of Animal Reproduction
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• v.23 no.3
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• pp.221-227
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• 1999

Successful cryopreservation of bovine immature oocytes can increase availably of oocytes for the in vitro fertilization or nuclear transfer. However, it was not reported successful development to the blastocyst stage following in vitro fertilization of cryopreserved bovine immature oocytes. The objective of this study was to determine the incidence of survival, meiotic maturation, fertilization and in vitro development of cryopreserved bovine immature by ultra rapid cooling methods. The oocytes were adversely affected by brief exposure to EFS40 solution in electron microscope grids and plunged directly into liquid nitrogen. After such ultra-rapid cooled immature oocytes were warmed, 78% of oocytes were matured to the metaphase II stage, 50% of oocytes were fertilized after insemination, and 5% of oocytes were developed to the blastocyst stage. Different sodium concentration of sodium ion in the freezing medium did not affect survival, maturation, fertilization and in vitro development of cryopreserved oocytes. These results suggested that immature bovine oocytes can be cryopreserved by ultra-rapid cooling methods.

• Reproductive and Developmental Biology
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• v.35 no.3
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• pp.197-201
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• 2011

This study was conducted to investigate the effects of donor cell treatments on the production of transgenic cloned piglets. Ear fibroblast cell obtained from NIH MHC Inbred minipig was used as control. The GalT knock-out/CD45 knock-in (GalT/CD46) transgenic cell lines were established and used as donor cells. The reconstructed GalT/CD46 embryos were surgically transferred into oviduct of naturally cycling surrogate sows (Landrace ${\times}$ Yorkshire) on the second day of standing estrus. Unlike control (1.2 kV/cm, 75.4%), the fusion rate of the GalT/CDl6 donor cells was significantly higher in 1.5 kV/cm, (84.5%) than that of 1.25 kV/cm, (20.2%) (p<0.01). When the number of the transferred embryos were more than 129, the pregnancy and delivery rates were increased to 13/20 (65%) and 5/20 (25%) compared to less then 100 group [1/6 (16.7%) and 0/6 (0%)], respectively. To analyze the effect of donor cell culture condition on pregnancy and delivery rates, the GalT/CD46 donor cells were cultured with DMEM or serum reduced medium. In serum reduced medium group, the pregnancy and delivery rates were improved to 8/12 (66.7%) and 5/12 (41.7%) compared to DMEM group [3/7 (42.9%) and 0/7 (0%)], respectively. In conclusion, it can be postulated that an appropriate fusion condition and culture system is essential factors to increase the efficiency of the production of transgenic cloned piglets.

• Korean Journal of Environmental Agriculture
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• v.17 no.3
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• pp.215-219
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• 1998

This experiment was conducted to investigate effects of NaCl concentration on photosynthetic rate, free proline content and ion content in tobacco. As NaCl concentration was increased growth was retarded. The decrease growth characteristics(shoot/root ratio was 2.0) at 90mM NaCl indicated that this concentration could be a limiting level. As NaCl concentration was increased photosynthetic rate, transpiration rate, and water use efficiency were decreased. Photosynthetic rate was highly decreased at 60mM NaCl. There was no significant difference between transpiration rate and water use efficiency. Leaf water potential was decreased as NaCl concentration was increased, in that twice lower at 30mM than that of control and drop largely at 120mM NaCl. Free proline content was increased as NaCl increased until 120mM NaCl and drop at 150mM NaCl. The $Ca^{2+}$, $Mg^{2+}$, and $K^+$ contents were increased until NaCl concentration was 120mM. The $Na^{2+}$ content was slowly increased as NaCl concentration increased until 120mM NaCl, and largely increased at 150mM NaCl. There was no significant difference between $Cl^-$ and NaCl treatments except 30 mM NaCl in which $Cl^-$ content was higher than that of control. As NaCl concentration was increased $K^+/Na^+$ ratio was decreased. The negative correlation between $K^+$ and $Na^+$, and positive correlation between $K^+/Na^+$ and protein content were found.

• The Journal of the Korean Society for Microbiology
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• v.21 no.3
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• pp.399-406
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• 1986

This study was performed to establish human plasmacytoma cell line as the partner cells for producing human hybridoma. Bone marrow cells from a multiple myeloma patient from Seoul National University Hospital, Korea were cultured and established as the cell line, named as HMC-BM4. HMC-BM4 cells were cultivated in RPMI 1640 media containing 8-azaguanine(8-AG; gradually increasing concentration from $1\;{\mu}g/ml$ to $20\;{\mu}g/ml$). 8-AG resistant cells were collected and cloned by limiting dilution. Each clone was divided and tested to die in hypoxanthine, aminopterine and thymidine (HAT) selection media. Finally one clone was selected and named as HMC-AR, which was sensitive to HAT selection media. HMC-AR cells showed typical morphology of plamacytoma in Wright staining. No cell formed the rosette with sheep erythrocytes. Surface membrane $\mu$ heavy chain was detected in 20% of HMC-AR cells and cytoplasmic $\mu$ heavy chain in 90% of them by direct immunofluorescent staining. Ia-like antigen was found in 90% of HMC-AR cells by indirect immunofluorescent staining using anti-Ia-like antigen monoclonal antibody, 1BD9-2. And about $1.0\;{\mu}g/ml$ of human $\mu$ heavy chain was detected in the 3-day culture supernatant of HMC-AR cells. 88% of cells contained 46 chromosomes. Mycoplasma was not detected in HMC-AR cells by Hoechst 33258 staining. This cell line would be used for making hybridomas secreting human monoclonal antibody.