• KSBB Journal
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• v.9 no.2
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• pp.191-199
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• 1994

Protoplasts of both strains were produced by lysozyme digestion at $30^{\circ}C$ for 180min. Both strains were treated with $40{\mu}g$/ml of lysozyme in 30mM Tris-HCl buffer(pH 7.5) containing 10% sucrose at the late logarithmic growth phase. It was found that the efficiency of protoplast formation was high at $30^{\circ}C$ and pH 7.5 by measuring the decrease in absorbance. Optimum concentrations of sucrose $Ca^{2+}, \;Mg^{2+}$ for protoplast formation were determined to be 15%, 20mM and 6mM, respectively. Hydrolysis of cell wall and protoplast formation efficiency for L. plantarum showed better results than those for Leu. mesenteroides. The resistances to antibiotics erythromycin and chloramphenicols were chosen as the selection marker for the fusant between L. plantarum and Leu. mesenteroides. Production phase of protoplast in Leu. mesenteroides was also compared with L. plantarum in this paper.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.10
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• pp.1407-1415
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• 2016

Isolation and culture of spermatogonial stem cells (SSCs) are attractive for production of genetic modified offspring. In the present study, buffalo spermatogonial stem-like cells were isolated, cultured and expression pattern of different germ cell marker genes were determined. To recover spermatogonia, testes from age 3 to 7 months of buffalo were decapsulated, and seminiferous tubules were enzymatically dissociated. Two types of cells, immature sertoli cell and type A spermatogonia were observed in buffalo testes in this stage. Germ cell marker genes, OCT3/4 (Pou5f1), THY-1, c-kit, PGP9.5 (UCHL-1) and Dolichos biflorus agglutinin, were determined to be expressed both in mRNA and protein level by reverse transcription polymerase chain reaction and immunostaining in buffalo testes and buffalo spermatogonial stem-like cells, respectively. In the following, when the isolated buffalo buffalo spermatogonial stem-like cells were cultured in the medium supplemented 2.5% fetal bovine serum and 40 ng/mL glial cell-derived neurotrophic factor medium, SSCs proliferation efficiency and colony number were significantly improved than those of other groups (p<0.05). These findings may help in isolation and establishing long term in vitro culture system for buffalo spermatogonial stem-like cells, and accelerating the generation of genetic modified buffaloes.

• Journal of the Institute of Electronics Engineers of Korea CI
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• v.49 no.4
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• pp.71-81
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• 2012

In order to support simple authoring an application of augmented reality for diverse users, the process for the object registration comprised of tracking and recognizing the object should be accomplished intuitively and simple. Although many 3D reconstruction methods to be applied to the object registration have been developed, the methods have not beyond the experimental level yet. In this paper, we proposed a novel marker-based tangible interfaces for various users to manipulate the object with intuitive and simple approaches during an authoring applications fo augmented reality. The proposed method make use of marker as intuitive interface to obtain 3D geometric information of 3D reconstruction. 3D geometric information of an object surface is acquired by touching the object directly with the proposed tangible interfaces. The tangible interfaces not only support 3D reconstruction for graphical modeling but also offer features information which is used for augmented reality. Finally, we verify efficiency of the proposed method with demonstration of an augmented reality application using the proposed method.

• Journal of Biomedical Engineering Research
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• v.44 no.1
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• pp.92-98
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• 2023

Purpose: To propose a new infusion rate monitoring technique based on the 2D image marker tacking to improve patient safety by preventing syringe pump-related medication accidents due to decreased infusion rate control accuracy. Materials and Methods: The infusion rate of the syringe pump and drug residue in the pump-equipped syringe were monitored in real time by tracking the movement of the 2D image markers attached to the syringe pump. Results: The error rate between the set and the estimated infusion rates was 1.03, 0.66, 1.95, 0.23, and 1.05% when the infusion rate setting was 10, 20, 30, 40, and 50 mL/H, respectively. In addition, the error rate between the actual and the estimated drug residues was 1.04, 0.47, 0.60, 3.66, and 0.00% when the infusion rate setting was 10, 20, 30, 40, and 50 mL/H, respectively. Conclusion: Experimental results demonstrated that the proposed technique can increase the efficiency of the safety management system for seriously ill inpatients by decreasing a possibility of syringe pump-related medication accidents in hospitals.

• The Plant Pathology Journal
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• v.39 no.5
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• pp.494-503
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• 2023

Xanthomonas campestris pv. campestris (Xcc) is a plant pathogen of Brassica crops that causes black rot disease throughout the world. At present, 11 physiological races of Xcc (races 1-11) have been reported. The conventional method of using differential cultivars for Xcc race detection is not accurate and it is laborious and time-consuming. Therefore, the development of specific molecular markers has been used as a substitute tool because it offers an accurate and reliable result, particularly a quick diagnosis of Xcc races. Previously, our laboratory has successfully developed race-specific molecular markers for Xcc races 1-6. In this study, specific molecular markers to identify Xcc race 7 have been developed. In the course of study, whole genome sequences of several Xcc races, X. campestris pv. incanae, X. campestris pv. raphani, and X. campestris pv. vesicatoria were aligned to identify variable regions like sequence-characterized amplified regions and insertions and deletions specific to race 7. Primer pairs were designed targeting these regions and validated against 22 samples. The polymerase chain reaction analysis revealed that three primer pairs specifically amplified the DNA fragment corresponding to race 7. The obtained finding clearly demonstrates the efficiency of the newly developed markers in accurately detecting Xcc race 7 among the other races. These results indicated that the newly developed marker can successfully and rapidly detect Xcc race 7 from other races. This study represents the first report on the successful development of specific molecular markers for Xcc race 7.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.56 no.1
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• pp.124-132
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• 2023

This study aimed to cross between Korean and Japanese pure lines of Neopyropia strains to establish cross breeding technology and identify a superior variety that harbors the strength of both parents. Four crossing combinations were tried using three methods, resulting in 1,476 single conchocelis colonies. The three co-dominant Cleaved Amplified Polymorphic Sequence (CAPS) markers (EF-1α/Mse I, TOP2/Mse I, car A/ApaL I) were used to distinguish heterozygotic sporophytes and their maternal lines obtained from the inter and intraspecific cross-fertilization within the wild type of Neopyropia strains. Of the 1,476 colonies, 26.9% (218) were heterozygotes obtained from the nuclear CAPS markers. Their maternal line was clearly confirmed using organelle CAPS marker and chimeric thallus was obtained from crossing experiment of Japanese N. yezoensis (♀) and Korean N. yezoensis (♂). The use of CAPS markers improved the efficiency of crossbreeding by quickly screening heterozygotes and maternal lines in the conchocelis phase, which otherwise required pigmentation mutants as genetic markers.

• Journal of Life Science
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• v.26 no.12
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• pp.1355-1359
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• 2016

Human bone marrow mesenchymal stem cells (hBM-MSCs) can differentiate into various cell types including osteoblasts, adipocytes, chondrocytes, and myocytes. Previous studies, including our own, have shown that MSCs can also differentiate into neuron-like cells. However, their rate of neuronal differentiation is not sufficient for application to stem cell therapy, which requires well-defined cell types. For this purpose, we first examined the expression of neuronal lineage markers (GFAP, MAP-2, KCNH1, Nestin, NF-M, and Tuj-1) by real-time PCR, western blot, and immunocytochemical staining. The expressions of the astrocyte marker GFAP and neuronal markers NF-M and Tuj-1 increased in neuronal differentiated MSCs (dMSCs). To improve the neuronal differentiation efficiency, PDE4, an important signaling intermediator in the progression of neuronal differentiation, was modulated using well-known inhibitors such as rolipram or resveratrol and then differentiated into neuronal cells (Roli- or RSV-dMSCs). The expressions of NF-M, Tuj-1 were increased while that of GFAP decreased in Roli- and RSV-dMSCs, which were examined by real-time PCR, western blot, and immunocytochemical staining. From these experiments, we have found that the neuronal differentiation efficiency can be ameliorated by the modulation of PDE4 activity.

• Animal Bioscience
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• v.37 no.5
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• pp.929-943
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• 2024

Objective: The present study was executed to explore the molecular mechanism of fibroblast growth factor 10 (FGF10) gene in bovine adipogenesis. Methods: The bovine FGF10 gene was overexpressed through Ad-FGF10 or inhibited through siFGF10 and their negative control (NC) in bovine adipocytes, and the multiplicity of infection, transfection efficiency, interference efficiency were evaluated through quantitative real-time polymerase chain reaction, western blotting and fluorescence microscopy. The lipid droplets, triglycerides (TG) content and the expression levels of adipogenic marker genes were measured during preadipocytes differentiation. The differentially expressed genes were explored through deep RNA sequencing. Results: The highest mRNA level was found in omasum, subcutaneous fat, and intramuscular fat. Moreover, the highest mRNA level was found in adipocytes at day 4 of differentiation. The results of red-oil o staining showed that overexpression (Ad-FGF10) of the FGF10 gene significantly (p<0.05) reduced the lipid droplets and TG content, and their down-regulation (siFGF10) increased the measurement of lipid droplets and TG in differentiated bovine adipocytes. Furthermore, the overexpression of the FGF10 gene down regulated the mRNA levels of adipogenic marker genes such as CCAAT enhancer binding protein alpha (C/EBPα), fatty acid binding protein (FABP4), peroxisome proliferator-activated receptor-γ (PPARγ), lipoprotein lipase (LPL), and Fas cell surface death receptor (FAS), similarly, down-regulation of the FGF10 gene enriched the mRNA levels of C/EBPα, PPARγ, FABP4, and LPL genes (p<0.01). Additionally, the protein levels of PPARγ and FABP4 were reduced (p<0.05) in adipocytes infected with Ad-FGF10 gene and enriched in adipocytes transfected with siFGF10. Moreover, a total of 1,774 differentially expressed genes (DEGs) including 157 up regulated and 1,617 down regulated genes were explored in adipocytes infected with Ad-FGF10 or Ad-NC through deep RNA-sequencing. The top Kyoto encyclopedia of genes and genomes pathways regulated through DEGs were the PPAR signaling pathway, cell cycle, base excision repair, DNA replication, apoptosis, and regulation of lipolysis in adipocytes. Conclusion: Therefore, we can conclude that the FGF10 gene is a negative regulator of bovine adipogenesis and could be used as a candidate gene in marker-assisted selection.

• Journal of Plant Biotechnology
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• v.46 no.4
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• pp.323-330
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• 2019

The high expression level of industrial and metabolically important proteins in plants can be achieved by plastid transformation. The CaIA vector, a Capsicum-specific vector harboring aadA (spectinomycin resistance), is a selectable marker controlled by the PsbA promoter, and the terminator is flanked by the trnA and trnI regions of the inverted repeat (IR) region of the plastid. The CaIA vector can introduce foreign genes into the IR region of the plastid genome. The biolistic method was used for chloroplast transformation in Scoparia dulcis with leaf explants followed by antibiotic selection on regeneration medium. Transplastomes were successfully screened, and the transformation efficiency of 3 transgenic lines from 25 bombarded leaf explants was determined. Transplastomic lines were evaluated by PCR and Southern blotting for the confirmation of aadA insertion and its integration into the chloroplast genome. Seeds collected from transplastomes were analyzed on spectinomycin medium with wild types to determine genetic stability. The increased chloroplast transformation efficiency (3 transplastomic lines from 25 bombarded explants) would be useful for expressing therapeutically and industrially important genes in Scoparia dulcis L.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.4
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• pp.532-537
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• 2005

Increasing of body weight has been one of the important economic factors in the poultry industry. Insulin-like growth factor (IGF)-I is a polypeptide that serves to regulate muscle development and body growth. Moreover, IGF-I is related to feed efficiency. However, there are few studies regarding the regulatory roles of chicken IGF-I/-II compared with that of mammals. Especially, the Korean Native Ogol Chicken (KNOC) has a lean body growth and its body weight is generally lighter than the broiler chicken. Therefore, this study was conducted to investigate associations among serum IGF-I/-II concentration, feed efficiency, and body growth in KNOC. The body weight and feed intake of KNOC were recorded from 20 to 36 weeks at 2 weeks intervals, and blood was taken every 2 weeks. Serum IGF-I/-II were measured by RIA. Chickens were divided into two groups, high and low serum IGF-I concentration. Generally, feed efficiency and growth performance (body weight and weight gain) in the high serum IGF-I group were higher than those of the low group during the experimental period. In particular, the body weight of the IGF-I high group were significantly different from those of the IGF-I low group at 34 and 36 weeks, respectively (p<0.05). Moreover, body weight, weight gain, and feed efficiency had a significant correlation with serum IGF-I at several weeks (p<0.05 and p<0.01). These results show that IGF-I plays an important role in body growth and suggests a possibility that serum IGF-I could be used as a selection marker for body growth in KNOC.