• Journal of Animal Science and Technology
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• v.46 no.5
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• pp.879-886
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• 2004

Effects of culture mediwn supplements and osmotic stress treatment on embryogenic callus induction and somatic embryogenesis were investigated in order to optimize tissue culture conditions of alfalfa(Medicago sativa L.). SH mediwn containing 5mgIL 2,4-D and 0.2mgIL kinetin was optimal for embryogenic callus induction from cotyledon tissue of alfalfa. Somatic embryos were formed when the embryogenic callus was cultured on SH mediwn supplemented with ImgIL 2,4-D and 2mgIL BA. Supplementation of 5mM L-proline and IgIL casein hydrolysate into the regeneration mediwn further increased plant regeneration frequency. Osmotic stress treatment of callus appeared to improve the frequency of somatic embryo formation, but the frequency of somatic embryo formation differed by the osmotic stress treatment using different osmotic stressors. The highest plant regeneration frequency of 30.7% was observed when embryogenic callus was treated with 0.7M sucrose for 18h. Efficient regeneration system established in this study will be useful for molecular breeding of alfalfa through genetic transformation.

• Plant Biotechnology Reports
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• v.4 no.4
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• pp.253-260
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• 2010

Plant secondary metabolites have always been a focus of study due to their important roles in human medicine and nutrition. We transferred the isoflavone synthase (IFS) gene into soybean [Glycine max (L.) Merr.] using the Agrobacterium-mediated transformation method in an attempt to produce transformed soybean plants which produced increased levels of the secondary metabolite, isoflavone. Although the trial to produce transgenic plant failed due to unestablished hygromycin selection, transformed callus cell lines were obtained. The induction rate and degree of callus were similar among the three cultivars tested, but light illumination positively influenced the frequency of callus formation, resulting in a callus induction rate of 74% for Kwangan, 67% for Sojin, and 73% for Duyou. Following seven to eight subcultures on selection media, the isoflavone content of the transformed callus lines were analyzed by high-performance liquid chromatography. The total amount of isoflavone in the transformed callus cell lines was three- to sixfold higher than that in control callus or seeds. Given the many positive effects of isoflavone on human health, it may be possible to adapt our transformed callus lines for industrialization through an alternative cell culture system to produce high concentrations of isoflavones.

• Journal of Sensor Science and Technology
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• v.6 no.6
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• pp.500-507
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• 1997

For fine control of operating temperature of shape memory alloy, we investigated the effect of thermal teratment of shape memory alloy with the impurity kind. The martensitic transformation temperature in a Cu-17.25Zn-15Al and Cu-17.25Zn-15Al-1Ag/Fe was measured using electrical resistivity as a function of quenching temperature. Order-disorder phase transition temperatures in parent phase were measured and kind of transition were distinguised by DSC(differential scanning calorimeter) with heating rate variation. And structual changes were studied with XRD. For the Cu-17.25Zn-15Al shape memory alloy, the order-disorder phase transition temperature, $T_{B2}$ and $T_{L21}$ was 809K and 610K and for the Cu-17.25Zn-15Al-1Ag and Cu-17.25Zn-15Al-1Fe specimen $T_{B2}$ and $T_{L21}$ was 794K and 610K, and 803K and 613K, respectively. In all the specimens, quenching from near $T_{B2}$ leads to an increase in martensitic temperature, whereas quenching from near $T_{L21}$ leads to an decrease in martensitic temperature.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.5
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• pp.587-594
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• 2000

At present embryonic stem (ES) cells with confirmed pluripotential properties are only available in the mouse. Recently, we were able to isolate, culture and genetically transform primordial germ cell (PGC)-derived cells from pig embryos and demonstrate their ability to contribute to chimera development in the pig. In order to determine whether the system we developed could be used to isolate embryonic germ (EG) cells from other mammalian species, we placed isolated PGCs from cattle, goats, rabbits and rats in culture. Briefly, PGCs were isolated from fetuses of cow (day 30-50), goat (day 25), rabbit (day 15-18) and rat (day 11-12), and plated on STO feeder cells in Dulbecco's modified Eagle's medium (DMEM): Ham's F10 medium (1:1) supplemented with 0.01 mM nonessential amino acids, 2 mM L-glutamine, 0.1 mM $\beta$ - mercaptoethnol, soluble recombinant human stem cell factor (SCF; 40ng/ml), human basic fibroblast growth factor (bFGF; 20ng/ml) and human leukemia inhibitory factor (LIF; 20ng/ml). For maintenance of the cells, colonies were passed to fresh feeders every 7-10 days. In all species tested, we were able to obtain and maintain colonies with ES-like morphology. Their developmental potential was tested by alkaline phosphatase (AP) staining and in vitro differentiation assay. For genetic transformation, cells were electroporated with a construct containing the green fluorescent protein (GFP) under the control of the cytomegalovirus (CMV) promoter. GFP-expressing colonies were detected in cattle, rabbits and rats. These results suggest that PGC-derived cells from cattle, goats, rabbits and rats can be isolated, cultured, and genetically transformed, and provide the basis for analyzing their developmental potential and their possible use for the precise genetic modification of these species.

• Asian-Australasian Journal of Animal Sciences
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• v.12 no.1
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• pp.22-27
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• 1999

Bovine oocytes with compact and complete cumulus cells were cultured in 6 groups for up to 24h in TCM199 buffered with 25 mmol/1 HEPES and supplemented with 10% FCS, 1 mg/ml $17{\beta}$-estradiol, 20 IU/ml hCG. Half of the oocytes at each group cultured in the presence of $25{\mu}g/ml$ cycloheximide at different times during maturation (0, 6, 12, 18, 20, 22 h) were fixed at 24 h of maturation to examine the nuclear progression. The rests of them were inseminated with frozen-thawed spermatozoa in medium BO with 10 mg/ml BSA and 10 mg/ml heparin and fixed after additional 18-20 h culture to evaluate the sperm head transformation. When a protein synthesis inhibitor was added at the onset of the maturation, the oocytes were prevented to proceed GVBD. A few of the oocytes (16%) were able to be penetrated and sperm head decondensation was inhibited either. Addition of cycloheximide after 6-12 h of culture resulted in an increasing percentage of GVBCD (more than 80%), but the oocytes became arrested in M-I (69.2%). More than half of the oocytes was penetrated with a decondensing sperm head. Formation of male pronucleus was first obtained at 12 h of culture in the presence of cycloheximide. When cycloheximide was added from 18 h of culture onwards, nuclear progression to M-II was increasingly restored (80.4-85.5%). The proportion of male and female pronuclear formation increased from 17.9% to 46.2%. It is concluded that protein synthesis is necessary not only for GVBD and development from M-I to M-II, but also for sperm head decendensation and male pronuclear formation in bovine oocytes.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.148.2-149
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• 2003

Cucumber fruit mottle mosaic tobamovirus (CFMMV) causes severe mosaic symptoms with yellow mottling on leaves and fruits, and occasionally severe wilting of cucumber plants. No genetic source of resistance against this virus has been identified. The genes coding for the coat protein or the putative 54-kDa replicase were cloned into binary vectors under control of the SVBV promoter. Agrobacterium-mediated transformation was peformed on cotyledon explants of a parthenocarpic cucumber cultivar with superior competence for transformation. R1 seedlings were evaluated for resistance to CFMMV infection by lack of symptom expression, back inoculation on an alternative host and ELISA. From a total of 14 replicase-containing R1 lines, 8 exhibited immunity, while only 3 resistant lines were found among a total of 9 CP-containing lines. Line 144 homozygous for the 54-kDa replicase was selected for further resistance analysis. Line 144 was immune to CFMMV infection by mechanical and graft inoculation, or by root infection following planting in CFMMV-contaminated soil. Additionally, line 144 showed delay of symptom appearance following infection by other cucurbit-infecting tobamoviruses. Infection of line 144 plants with various potyviruses and cucumber mosaic cucumovirus did not break the resistance to CFMMV. The mechanism of resistance of line 144 appears to be RNA-mediated, however the means is apparently different from the gene silencing phenomenon. Homozygote line 144 cucumber as rootstock demonstrated for the first time protection of a non-transformed scion from soil inoculation with a soil borne pathogen, CFMMV.

• Journal of Plant Biotechnology
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• v.42 no.4
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• pp.396-400
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• 2015

To investigate the optimal conditions for shoot organogenesis in Astragalus sinicus L., hypocotyl explants were cultured in Murashige & Skoog's (MS) medium supplemented with 0.1, 1.0, 2.0, or 4.0 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D) for 6 weeks. 2,4-D concentration significantly effected morphogenesis: some produced calli with adventitious shoots and roots, some produced calli with adventitious roots, some produced only calli, and some produced deep-brownish calli with roots. The formation of calli with shoots and/or roots was observed at lower levels of 2,4-D, whereas calli without shoots or with deep-brownish roots were formed after treatment with higher levels of 2,4-D. Also, a shoot organogenesis ability of callus clones was observed after treatment with medium with 0.1 or 1.0 mg/L 2,4-D grown in MS medium with combinations of benzyl adenine (BA) and 2,4-D for 4 weeks. Medium with a combination of BA and 2,4-D was effective for shoot formation, whereas root organogenesis from calli decreased. The greatest amount of shoot formation was obtained when calli were cultured in MS medium containing 1.0 mg/L 2,4-D and 0.5 mg/L BA. Upon shoot transfer into 1/2 MS basal medium, plantlets developed, and the plantlets grew well in soil in a greenhouse.

• Journal of Sericultural and Entomological Science
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• v.49 no.2
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• pp.37-42
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• 2007

Lactoferrin, an ion-binding 80-kDa glycoprotein, has been suggested to have many biologic activities, such as facilitating ion absorption and having antimicrobial and anti-inflammatory effects. Several of these activities are likely to only be facilitated by human lactoferrin because they depend on the binding of human lactoferrin to specific receptor. To produce recombinant human lactoferrin to animal foods using transgenic silkworm, Bombyx mori L, we have cloned and sequenced the cDNA encoding for a human lactoferrin (HLf) from the mRNA in mammary tumor line (GI-101). As a result, the 2.5-kb fragment of HLf gene was cloned with pGEM-T vector and then this fragment was sequenced. In the nucleotide sequence analysis, single open reading frame of the 2,136-bp encoding for a polypeptide of 712 amino acid residues was detected. On the other hand, we constructed a recombinant plasmid(pPT-HLf), containing human lactoferrin gene for germ line transformation of the silkworm using a piggyBac transposon-derived vector. A nonautonomous helper plasmid encodes the piggyBac transposase. Approximately 6.7% of individuals in the G0 silkworms expressed green fluorescent protein (GFP). PCR analyses of GFP-positive silkworms (G0 and G1) revealed that independent insertions occurred frequently. Furthermore, Western blot analysis showed that the recombinant HLf expressed in hemolymph has the same molecular weight (80 kDa) as a native protein. On the basis of these experiments, expression of HLf in next generation of transgenic silkworm is now in process.

• Korean Journal of Plant Tissue Culture
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• v.26 no.4
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• pp.281-287
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• 1999

Codonopsis lanceolata ("Deoduck" in Korea) is a perennial herb, and belongs to family, Campanulaceae. Its taproot is used a good source of a wild vegetable as well as an herbaceous medicine. In this study, to develop a bialaphos-resistant transgenic Codonopsis, seed germination mechanism and somatic embryogenesis of the plant were investigated, and Agrobacterium-mediated transformation with bar gene encoding phosphinothricin acetyltransferase (PAT) was performed. Attempt were made to regenerate plant from cells via somatic embryogenesis. When the cotyledons, nodes and leaf disks were cultured on MS medium containing 2,4-D and zeatin, embryogenic calli were induced. Upon transferring the somatic embryos to N6 solid medium without plant growth regulators, they developed into plantlets under continuous illumination. All plants were dead on MS basal medium containing 10 mg/L phosphinothricin (PPT) and Basta, respectively. The explants did not produce calli in the medium containing 200 mg/L kanamycin. The explants were cocultured with Agrobacterium tumefaciens for 2 days, and transformants were selected in MS basal medium containing 1.0 mg/L 2,4-D, 100 mg/L kanamycin and 500 mg/L carbenicillin. After the selection, embryogenic calli were induced and then somatic embryos were produced by subsequent subculturing. The somatic embryos were germiated on N6 basal medium containing 200 mg/L kanamycin and 500 mg/L carbenicillin. PCR analysis showed that nptII and bar genes were introduced in the Deoduck transformants. After the confirmation of bar gene expression in RNA and protein level, the transgenic Deoduck will be used to study the genetics of filial generation with the herbicide control gene, bar.gene, bar.

• Journal of the Korean Society of Clothing and Textiles
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• v.15 no.3 s.39
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• pp.239-250
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• 1991

The purpose of this study was primarily aimed at the revealing changes in the aspects of women's clothing derivationed with the women's self-consciousness, liberation movement, women's suffrage and social participation from the mid-l9th century to the mid-20th century. According to the study, the main change of western women's costumes were eradication of corset, emergence of troussers and tailored suit, and transformation of sportwear. So that, women's liberation movement has an deep effect on leading up to a change tendency the functionalization and masculinization of women's costume.