• Journal of Embryo Transfer
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• v.15 no.2
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• pp.167-173
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• 2000

The principal objective of this study was to clone transgenic embryos in order to improve the efficiency of transgenic animal production by the combination of microinjection and nuclear transplantation techniques. Mature female New Zealand White rabbits were superovulated by eCG and hCG treatments, fllowed by natural mating. Zygotes were collected from the oviducts at 18∼22 h after hCG injection by flushing with D-PBS containing 5% fetal calf serum(FCS). Two to three picoliters of green fluorescent protein(GFP) gene wa microinjected into male pronucleus. The foreign gene-injected zygotes were cultured in TCM-199 or RD medium containing 10% FCS with a monolayer of rabbit oviductal epithelial cells in a 5% CO2 incubator. The morulae expressing GFP gene were selected and their blastomeres were separated for the use of nuclear donor. Following nuclear transplantation of fluorescence-positive morula stage blastomeres, 13 (21.3%) out of 61 fused oocytes developed to blastocyst stage and all of the cloned blastocysts expressed GFP. The results indicate that the screening of transgene in rabbit embryos by GFP detection could be a promisible method for the preselection of transgenic embryos. Also the cloning of preselected transgenic embryos by nuclear transplantatin could be efficiently applied to the multiple production of transgenic animals.

• The Korean Journal of Physiology and Pharmacology
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• v.23 no.6
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• pp.539-547
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• 2019

Anoctamin 5 (ANO5)/TMEM16E belongs to a member of the ANO/TMEM16 family member of anion channels. However, it is a matter of debate whether ANO5 functions as a genuine plasma membrane chloride channel. It has been recognized that mutations in the ANO5 gene cause many skeletal muscle diseases such as limb girdle muscular dystrophy type 2L (LGMD2L) and Miyoshi muscular dystrophy type 3 (MMD3) in human. However, the molecular mechanisms of the skeletal myopathies caused by ANO5 defects are poorly understood. To understand the role of ANO5 in skeletal muscle development and function, we silenced the ANO5 gene in C2C12 myoblasts and evaluated whether it impairs myogenesis and myotube function. ANO5 knockdown (ANO5-KD) by shRNA resulted in clustered or aggregated nuclei at the body of myotubes without affecting differentiation or myotube formation. Nuclear positioning defect of ANO5-KD myotubes was accompanied with reduced expression of Kif5b protein, a kinesin-related motor protein that controls nuclear transport during myogenesis. ANO5-KD impaired depolarization-induced $[Ca2^{+}]_i$ transient and reduced sarcoplasmic reticulum (SR) $Ca^{2+}$ storage. ANO5-KD resulted in reduced protein expression of the dihydropyridine receptor (DHPR) and SR $Ca^{2+}-ATPase$ subtype 1. In addition, ANO5-KD compromised co-localization between DHPR and ryanodine receptor subtype 1. It is concluded that ANO5-KD causes nuclear positioning defect by reduction of Kif5b expression, and compromises $Ca^{2+}$ signaling by downregulating the expression of DHPR and SERCA proteins.

• The Korean Journal of Nuclear Medicine Technology
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• v.22 no.2
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• pp.101-104
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• 2018

Purpose Alpha fetoprotein(AFP) is a fetal serum protein that increases in germ cell tumors derived from liver cancer or egg yolk. AFP test has been used for screening of liver cancer, determination of tumor stage, determination of therapeutic effect, and fetal congenital malformations. The purpose of this study was to compare the results of the four kits, identify the advantages and disadvantages of each kit, and select the appropriate kits for our laboratory. Materials and Methods Blood samples were obtained from 89 patients attending the Seoul national university hospital. Experiments were carried out in accordance with manufacturer's instructions of four companies(A, B, C, D). The precision, recovery, linearity, and sensitivity test were performed for each kit. Results In case of the precision within the measurement, the CV value of the C kit was less than 5% at the low, middle, and high concentrations. The A, B and D kit's the CV value was less than 5% at the concentrations except the low concentration. The recovery rates of the A, B, C, and D kits were $100{\pm}15%$, $100{\pm}30%$, $100{\pm}16%$ and $100{\pm}14%$, respectively. All kits showed good linearity. Sensitivity was measured as 0.5 IU/mL for A, 0.4 IU/mL for B, 0.98 IU/mL for C, and 0.3 IU/mL for D. Conclusion The CV values of the four kits were within 10%, and the correlation coefficients were close to 1 for $R^2=0.978$, $R^2=0.992$ and $R^2=0.8957$. As a result, they are clinically available. Therefore, each laboratory should select the appropriate kit for their experiment's environment.

• The Journal of Korean Society of Virology
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• v.26 no.1
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• pp.131-137
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• 1996

The sequences of p10 gene its promoter of Hyphantria cunea NPV were determined. According to the sequence analysis, the putative p10 gene ORF has 285 bp. The 5'-non-coding leader sequence of the p10 gene promoter contained the TATA box and the putative transcription initiation site TAAG motif. Poly (A) tail signals, AATAAA sequence was at site 65 base upstream from the 3' terminus. The deduced amino acid sequence of p10 protein was 95 with a predicted molecular weight of 10.26 kDa. In the p10 protein sequence, a hydrophobic region was present at the N-terminus of the protein, whereas the C-terminus was highly hydrophilic. The p10 protein of H. cunea NPV did not contain cysteine, histidine, trytophan, tryptophane, tyrosine, glutamine and asparagine residues.

• Microbiology and Biotechnology Letters
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• v.21 no.5
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• pp.406-413
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• 1993

This study was performed to produce monoclonal antibodies to the major polyhedral inclusion body (PIB) antigen of the occluded form of Hyphantria cunea nuclear polyhedrosis virus (HcNPV). PIB proteinS were purfied from the Spodoptera frugiperda cell infected with HcNPVs. Using the purified PIB protein, BALB/C mice were immunized 3 times with 2 weeks intervals. The spleen were removed and fused with Sp2/0-Ag14, mouse myeloma cells.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.7
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• pp.958-963
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• 2006

The effect of ginsenosides (GS) on germ cell proliferation was evaluated with a chicken ovarian germ-somatic cell coculture model and the mechanism involving protein kinase C (PKC) pathway was investigated. Ovarian cells were cultured in serum-free McCoy's 5A medium and challenged with GS alone or in combinations with PKC activator (phorbol 12-myristate 13-acetate, PMA) or inhibitor ($H_7$) for 48 h. The number of germ cells was counted and the proliferating cells were identified by immunocytochemistry of proliferating cell nuclear antigen (PCNA). Results showed that GS significantly increased germ cell proliferation and this stimulating effect was further increased by PMA, but inhibited by H7, in a dose-dependent manner. Moreover, GS-elevated PCNA expression and the PCNA -labeling index of germ cells displayed similar changes with the increased numbers of germ cells. These results indicated that GS stimulated proliferation of ovarian germ cells with involvement of the PKC-mediated system.

• Microbiology and Biotechnology Letters
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• v.21 no.5
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• pp.421-427
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• 1993

Some interleukin 6 (IL-60 transcription control factors were resported as the regulator of IL-6 expression. A nuclear protein bound to interleukin 1 (IL-1) responsive element in the IL-6 promoter region was named NF-IL6 (nuclear factor for IL-6). This NF-IL6 was known to be very imporant as a transcription factor for various immuno-protein as well as for IL-6. The human NF-IL6 genes were transfected into the mouse L cells under the metallothionein promoter (MT promoter) to establish a model system for the expression of foreign gene in the mammalian cell line.

• Journal of Radiopharmaceuticals and Molecular Probes
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• v.1 no.2
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• pp.95-97
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• 2015

The translocator protein (TSPO) is one of the important targets for Positron Emission Tomography (PET) imaging because it is associated with brain cancer, stroke, and neurodegeneration. Recently, a novel quinoline compound with high affinity agent for the translocator protein has been developed. In this highlight review, major studies for the quinoline compound are described.

• Journal of Radiopharmaceuticals and Molecular Probes
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• v.2 no.1
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• pp.37-42
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• 2016

The translocator protein (TSPO) has attracted scientist's attention for Positron Emission Tomography (PET) imaging due to correlation with brain cancer, stroke, and neurodegeneration. Recently, GE-180, a novel tricyclic derivative has been developed as a new high affinity agent for the TSPO and evaluated to confirm a possibility for the TSPO ligand. In this highlight review, several studies for the novel TSPO radiotracer are described.

• Korean Journal of Microbiology
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• v.7 no.1
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• pp.10-21
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• 1969

The effect of glucose and 2-thiobarbituric acid on the biosynthesis of cell constituents such as protein, carbohydrate, DNA, RNA, phospholipid and PCA-soluble phosphate compounds in Chlorella duing the life cycle was measured, and the changes in the content of these main cellular components of the algal cell were analyzed in connection with the nuclear and cytoplasmic divison. In the normal autotrophic synchronous culture the contents of protein, RNA, and DNA in the cell showed a chracteristic changes according to the progress of cell development, increasing more or less throughout all the life cycle. The synthesis of protein is more prominent in the division period nad that of DNA is more active in the ripening period, while the synthesis of RNA is more rapid in the growing and ripening periods than other developmental stages. The period of division cycle was little affected by glucose in the medium, although the synchrony of the growth and cellular division was disturbed and the n value increased. The cotents of protein, carbohydrate, RNA nad DNA of the cell were increased by the glucose treatment throughout all the life cycle. On the other hand, both of cellular growth and division were retarded severely and the n value was decreased by the 2-thiobarbituric acid treatment throughout all the life cycle. On the other hand, both of cellular growth and division were retarded severely and the n value was decreased by the 2-thiobarbituric acid treatment. The synthesis of protein, carbohydrate, DNA, RNA and phospholipid of the cell was also retarded by 2-thiobarbituric acid. In the autotrophic, mixotrophic and 2-thiobarbituric acid-treated cultures, each having different mode cytoplasmic division, a common general schema occurring in the cell during the life cycle may be drawn as follows. The ratio of RNA to protein attains maximum value in the $L_1$-cell stage prior to the nuclear division and thereafter decreases during the periods of ripening and division. The ratio of PCA-soluble phosphate compounds to protein increased from the begining of the culture to $L_4$-cell stage successively and thereafter decreased gradually during the division period, while the ratio of protein to DNA kept almost constant up to the division period and thereafter increased during the division period. Therefore, it is presumed that the increase in the ratio of RNA to protein is to be an inducer of nuclear division and that the cytoplasmic division is induced by the increase in the ratio of protein to DNA.