• Molecules and Cells
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• v.38 no.11
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• pp.950-958
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• 2015

BCS1L gene encodes mitochondrial protein and is a member of conserved AAA protein family. This gene is involved in the incorporation of Rieske FeS and Qcr10p into complex III of respiratory chain. In our previous study, AluYRa2-derived alternative transcript in rhesus monkey genome was identified. However, this transcript has not been reported in human genome. In present study, we conducted evolutionary analysis of AluYRa2-exonized transcript with various primate genomic DNAs and cDNAs from humans, rhesus monkeys, and crabeating monkeys. Remarkably, our results show that AluYRa2 element has only been integrated into genomes of Macaca species. This Macaca lineage-specific integration of AluYRa2 element led to exonization event in the first intron region of BCS1L gene by producing a conserved 3' splice site. Intriguingly, in rhesus and crabeating monkeys, more diverse transcript variants by alternative splicing (AS) events, including exon skipping and different 5' splice sites from humans, were identified. Alignment of amino acid sequences revealed that AluYRa2-exonized transcript has short N-terminal peptides. Therefore, AS events play a major role in the generation of various transcripts and proteins during primate evolution. In particular, lineage-specific integration of Alu elements and species-specific Alu-derived exonization events could be important sources of gene diversification in primates.

• Proceedings of the Korean Vacuum Society Conference
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• 2013.02a
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• pp.108-109
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• 2013

Mitochondria play key roles in the production of cell's energy. Their dominant function is the synthesis of adenosine 5'-triphosphate (ATP) from adenosine diphosphate (ADP) and phosphate (Pi) through the oxidative phosphorylation. Evaluation of drug-induced mitochondrial toxicity has become increasingly important since mitochondrial dysfunction has recently been implicated in numerous diseases including cancer and diabetes mellitus. Mitochondrial functions have been monitored via oxygen consumption, mitochondrial membrane potential, and more importantly via ATP synthesis since ATP synthesis is the most essential function of mitochondria. Various analytical methods have been employed to investigate ATP synthesis in mitochondria, including high performance liquid chromatography (HPLC), bioluminescence technique, and pH measurement. However, most of these methods are based on destructive analysis or indirect monitoring through the enzymatic reaction. Infrared absorption spectroscopy (IRAS) is one of the useful techniques for real-time, label-free, and direct monitoring of biological reactions [1,2]. However, the strong water absorption requires very short path length in the order of several micrometers. Transmission measurements with thin path length are not suitable for mitochondrial assays because solution handlings necessary for evaluating mitochondrial toxicity, such as rapid mixing of drugs and oxygen supply, are difficult in such a narrow space. On the other hand, IRAS in the multiple internal reflection (MIR) geometry provides an ideal optical configuration to combine solution handling and aqueous-phase measurement. We have recently reportedon a real-time monitoring of drug-induced necrotic and apoptotic cell death using MIR-IRAS [3,4]. Clear discrimination between viable and damaged cells has been demonstrated, showing a promise as a label-free and real-time detection for cell-based assays. In the present study, we have applied our MIR-IRAS system to mitochondria-based assays by monitoring ATP synthesis in isolated mitochondria from rat livers. Mitochondrial ATP synthesis and hydrolysis were in situ monitored with MIR-IRAS, while dissolved oxygen level and solution pH were simultaneously monitored with O2 and pH electrodes, respectively. It is demonstrated that ATP synthesis and hydrolysis can be monitored by the IR spectral changes in phosphate groups in adenine nucleotides and MIR-IRAS is useful for evaluating time-dependent drug effects of mitochondrial toxicants.

• The Korean Journal of Zoology
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• v.31 no.3
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• pp.236-242
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• 1988

Mitochondrial DNAs of two Mdh allelotypes of the dark chub, Z. temmincki inhabiting in Korean fresh water, were analysed. Samples of each type were collected from four populations, and the fragment patterns for mtdNA of each type were explained from 7 of the eleven restriction enzymes with hexanucleotide recognition site. Genome size was approximately 16.7 kilobases. The highly typical mtdNA fragments of each type were discovered in digestion profiles produced by Eco RI and Pst I enzyrnes. The comparisons of restriction fragment patterns and relative digestion maps permitted the estimation of fragment homology (F) and nucleotide sequence divergence(p). Between the two identical types, sequence divergence(p) was 0.128(MS), and 0.045(MM), ; between the two different types, 0.195 (range 0.177-0.226). These result may provide a distinct difference more than the value derived from allozyrne analysis, and a powerful new molecular approach for assessing genetic-evolutionary relationship among fishes.

• Journal of Science and Technology Studies
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• v.1 no.2 s.2
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• pp.423-438
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• 2001

The purpose of this study is to understand how a teacher's teaching can be changed while he or she teaches the same contents in different classes. The qualitative research method was used in this study. Data were collected from classroom observations, several in-depth interviews, and stimulated-recall interviews after each class. All the data were transcribed and analyzed interpretively, and then, the results of the analysis were checked by each participating teacher. The results are as follows: First, changes appeared in each class in terms of the teaching items, tools, sequence, and time, even though the same teacher taught the same contents. It showed that the teacher's teaching practice changed immediately and intuitively in class. Second, teachers tried to implement "exploratory teaching" or "move-testing teaching" to address the emerging problems during their teaching. They then reflected on and modified their own teaching. This type of change, which happened during the teaching practice, can be an example of "Reflection-in-practice." Thus, the results of this study can provide helpful insights into how teachers might adapt and reflect in their teaching. It suggests that teachers need to recognize their subconscious teaching changes and learn "Reflection-in-practice."

• Clinical and Experimental Reproductive Medicine
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• v.22 no.2
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• pp.191-201
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• 1995

Transcriptional activation of the embryonic genome initiates at 2-cell stage in mouse embryo and is characterized by the synthesis of TRC which is restricted to 2-cell stage. To investigate the roles of various protein kinases on the embryonic gene activation, the effects of protein kinase inhibitors on in vitro development and protein synthetic profiles of the early mouse embryos were examinded. None of ${\alpna}-amanitin$ which is a mRNA synthetic inhibitor, H8 which is a PKA inhibitor, and H7 which is a PKC inhibitor, affected on first cleavage of mouse 1-cell embryos in vitro. However, all of these drugs inhibited the second cleavage. When the drugs were removed following treatment for 6 hours, H8 or H7 treatment showed little inhibition on subsequent development of 1-cell embryos to 2-cell stage or further. In contrast, ${\alpna}-amanitin$ irreversibly inhibited the development of 1-cell embryos to 2-cell stage following removal of the drug. Genistein, a TPK inhibitor, inhibited both the first cleavage of 1-cell embryos and the second cleavage of 2-cell embryos, suggesting that TPK activity may be important during the early cleavages. All of the above four drugs inhibited TRC synthesis as shown by the fluorographic analysis of $[^{35}S]-Met$ labeled protein profiles. When late 1-cell embryos were treated with H7 and analyzed synthetic patterns of $[^{35}S]-Met$ labeled protein, the quantitative differences of protein synthesis on SDS-PAGE appeared on 77 kD and 33 kD region at $32{\sim}38$ hours post hCG. From these studies, transcriptional activation of embryonic genome is not essenting to the mouse 1-cell embryos to develop to 2-cell stage. Hawever, TPK activity is reguisite for both the first cleavage and second cleavage. Similarly, both PKC and PKA activities are required for the second cleavage of mouse embryos, but not for the first cleavage.

• Journal of Life Science
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• v.21 no.12
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• pp.1726-1731
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• 2011

Human Stem Cell Factor (hSCF) is a cytokine that binds to the c-Kit receptor and plays an important role in hematopoiesis, spermatogenesis, and melanogenesis. To produce the human Stem Cell Factor (hSCF) recombinant protein, we constructed a germline transgenic silkworm using the piggyback vector. The expression of the hSCF gene was driven by the Drosophila heat shock protein 70 (dHsp70) promoter. 3XP3 promotor-driven EGFP was used as a marker which allowed us to rapidly distinguish the transgenic silkworm. A mixture of the donor and helper vector was micro-injected into 1,020 eggs of bivoltin silkworms, Keomokjam. We obtained approximately 22 G1 broods that were EGFP-positive. The expression of the hSCF gene in the transgenic silkworm was analyzed by SDS-PAGE and immunoblotting. Also, analysis of insertion sites into the silkworm genome using inverse PCR showed that exogenous DNA was inserted into the transgenic silkworm genome. These results show that successfully constructed transgenic silkworm expresses the hSCF recombinant protein.

• Journal of Animal Science and Technology
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• v.46 no.2
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• pp.235-242
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• 2004

A system for the production of transgenic plants has been developed for Italian ryegrass(Lolium mult리orum Lam.) via Agrobacterium-mediated transformation of embryogenic callus. Mature seed-derived calli were infected and co-cultured with Agrobacterium EHA101 carrying standard binary vector pIG121Hm encoding the hygromycin phosphotransferase(HPT), neomycin phosphotransferase II (NPTII) and intron-oontaining $\beta$g1ucuronidase( intron-GUS) genes in the T-DNA region. The effects of several factors on transformation and the expression of the GUS gene were investigated. Inclusion of 200${\mu}M$ acetosyringone(AS) in inoculation and co-cultivation media lead to a significant increase in stable transformation efficiency. Increasing Agrobacterium cell density up to 1.0 in $OD_{600}$ during infection increased transfonnation efficiency of embryogenic calli. The highest transfonnation efficiency was obtained when embryogenic calli were incoulated with Agrobacterium in the presence of 0.1% Tween20 and 200${\mu}M$ AS. Hygromycin resistant calli were developed into complete plants via somatic embryogenesis. GUS histochemical assay and PCR analysis of transgenic plants demonstrated that transgenes were integrated into the genome of Italian ryegrass.

• Korean Journal of Plant Resources
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• v.34 no.4
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• pp.278-286
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• 2021

Efficient in vitro regeneration system is essential for the successful crop breeding of soybean (Glycine max (L.) Merr.) using the new biotechnology. The genotype of donor plants strongly influences the establishment of tissue culture system. Therefore, the screening of genotypes with excellent tissue culture ability is very important for soybean genetic improvement. In this study, we report the tissue culture efficiency of 21 soybean cultivars belong to Korean soybean core-collection and two foreign cultivars (Jack and Maverick). The Kwangan, Anpyeong and Seonam are share close genetic relationship in 21 cultivars and these three cultivars were observed the high frequency of germination and regeneration. Furthermore, the high tissue culture abilities were also observed in the Williams 82 used in reference genome sequencing and the two foreign cultivars. The transformation of pBAtc:tRNA with bar gene was performed by Agrobacterium tumefaciens in the cultivars with high tissue culture ability. Transformation of the bar gene was identified by PCR analysis in Kwangan, Pungwon, Seonam, and Maverick. Our results provide useful information for the breeding of various soybean cultivars by plant biotechnology such as, genome editing.

• Korean Journal of Microbiology
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• v.55 no.2
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• pp.103-111
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• 2019

DNA methylation is involved in diverse processes in bacteria, including maintenance of genome integrity and regulation of gene expression. CcrM, the DNA methyltransferase conserved in Alphaproteobacterial species, carries out $N^6$-adenine or $N^4$-cytosine methyltransferase activities using S-adenosyl methionine as a co-substrate. Celeribacter marinus IMCC12053 from the Alphaproteobacterial group was isolated from a marine environment. Single molecule real-time sequencing method (SMRT) was used to detect the methylation patterns of C. marinus IMCC12053. Gibbs motif sampler program was used to observe the conversion of adenosine of 5'-GANTC-3' to $N^6$-methyladenosine and conversion of $N^4$-cytosine of 5'-GpC-3' to $N^4$-methylcytosine. Exocyclic DNA methyltransferase from the genome of strain IMCC12053 was chosen using phylogenetic analysis and $N^4$-cytosine methyltransferase was cloned. IPTG inducer was used to confirm the methylation activity of DNA methylase, and cloned into a pQE30 vector using dam-/dcm- E. coli as the expression host. The genomic DNA and the plasmid carrying methylase-encoding sequences were extracted and cleaved with restriction enzymes that were sensitive to methylation, to confirm the methylation activity. These methylases protected the restriction enzyme site once IPTG-induced methylases methylated the chromosome and plasmid, harboring the DNA methylase. In this study, cloned exocyclic DNA methylases were investigated for potential use as a novel type of GpC methylase for molecular biology and epigenetics.

• Journal of Plant Biotechnology
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• v.46 no.3
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• pp.180-188
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• 2019

Auxin plays a crucial regulatory role in plant growth and development processes. Three major classes of auxin-responsive transcription factors controlled by the Auxin/indole-3-acetic acid (Aux/IAA), Gretchen Hagen 3 (GH3), and small auxin up RNA (SAUR) genes regulate auxin signaling. Aux/IAA, in particular, encodes short-lived nuclear proteins that accumulate rapidly in response to auxin signaling. In this study, we isolated a PagAux/IAA1 gene from poplar (Populus alba ${\times}$ P. glandulosa) and investigated its expression characteristics. The PagAux/IAA1 cDNA codes for putative 200 amino acids polypeptide containing four conserved domains and two nuclear localization signals (NLSs). Utilizing Southern blot analysis, we confirmed that a single copy of the PagAux/IAA1 gene was present in the poplar genome. The expression of this gene is specific to leaves and flowers of the poplar. PagAux/IAA1 expressed in the early exponential growth phase of cell-cultured in suspension. PagAux/IAA1 expression level reduced in drought and salt stress conditions, and the presence of plant hormones such as abscisic acid. However, expression enhanced in cold stress, cambial cell division, and presence of plant hormones such as gibberellic acid and jasmonic acid. Thus, these results suggest that PagAux/IAA1 participates in cold stress response as well as developmental processes in the poplar.