• Journal of Life Science
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• v.27 no.6
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• pp.617-622
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• 2017

In this study, we sought to develop an effective cloning system by which human cytochrome $b_5$ (cyt $b_5$) is introduced and expressed in zebrafish. First, the 414 bp human cyt $b_5$ gene was amplified from RNA extracts of HeLa cells using RT-PCR, and the amplicon was subsequently sequenced to confirm that it was intact. Next, cyt $b_5$ was cloned into the pEGFP-N3 vector, which also encodes a fluorescent gene. One-cell stage zebrafish embryos were microinjected with the recombinant vector containing the cyt $b_5$ gene. Fluorescence microscopy confirmed high expression of the fluorescent gene in the injected fry compared to the non-fluorescent control fry. Finally, we extracted RNA from the injected fry and performed RT-PCR to determine whether the human cyt $b_5$ gene is expressed in the transgenic zebrafish. Sequencing analysis further confirmed that the cloned human cyt $b_5$ gene was intact. The transgenic zebrafish model produced in this study will be a useful tool to study therapeutic approaches to cure various diseases related to the deficiency of functional human cyt $b_5$ as well as tools for cloning useful genes in fish.

• YAKHAK HOEJI
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• v.31 no.6
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• pp.361-369
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• 1987

The four azo dyes such as Amaranth (FD & C Red No. 2), Tartrazine (FD & C Yellow No. 4), sunset Yellow (FD & C Yellow No. 5) and Allura red (FD & C Red No. 40) are currently employed as a food additives in Korea. In this study, the effects of these azo dyes on the hepatic microsomal mixed function oxidase systems in Rats. (i.e., Cyt. P-450, Cyt. b$_5$, NADPH cyt. c-reductase and azo reductase) were investigated. Furthermore, to determine the relationship among the electron transport systems, each level of azo reductase, Cyt. P-450 and NADPH cyt. c-reductase was measured upon the administration of phenobarbital (known as an inducer of Cyt. P-450), 3-methylcholanthrene (Known as an inducer of Cyt. P-448), CoCl$_2$ (inhibitor on Cyt. P-450) or $CCl_4$ (inhibitor on Cyt. P-450). The results of these studies are as follows; (1) The levels of Cyt. P-450 and Cyt. b$_5$ were decreased upon the administration of these azo dyes. (2) When the level of Cyt. P-450 was decreased, the azo reductase activity was also decreased. (3) These azo dyes did not show any significant effect on the level of NADPH cyt. c-reductase. (4) The administration of 3-methylcholanthrene resulted in the elevation of azo reductase activity. The 3-methylcholanthrene may be responsible for the induction of CO-insensitive electron transport system.

• Korean Journal of Ichthyology
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• v.18 no.1
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• pp.12-19
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• 2006

To examine the relationship of geological distribution and phylogenetic tree of O. latipes in the Far East, we analyzed cytochrome b (cyt b) gene in the mitochondrial genome. In this study we employed the entire sequence of cyt b of 53 samples collected from nine Korean locations and 117 cyt b data retrieved from the GenBank. From 170 Oryzias latipes cyt b sequence data, 142 different haplotypes were identified and phylogenetic relationship was reconstructed based on the dataset. According to the phylogeny, haplotypes were divided into three major haplogroups A, B and C, and their relationships were well correlated to their distributional patterns. Haplogroup A which is widely distribute in the southern part of Korea is separated in the geographical distribution from the haplogroup B which is found from China to the western part of Korea. Haplogroup C is only found in Japan.

• Journal of Microbiology and Biotechnology
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• v.23 no.1
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• pp.88-91
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• 2013

The Cyt1Aa protein of Bacillus thuringiensis subsp. israelensis is known to synergize mosquitocidal proteins of B. thuringiensis and Bacillus sphaericus strains. Cyt1Aa is highly lipophilic, and after binding in vivo to the midgut microvillar membrane serves as a "receptor" for mosquitocidal Cry proteins, which subsequently form cation channels that kill mosquito larvae. Here we report that Cyt1Aa can serve a similar function for lepidopteran-specific Cry proteins of B. thuringiensis in certain mosquito larvae. Engineering Cyt1Aa into the HD-1 isolate of B. thuringiensis subsp. kurstaki enhanced toxicity against $4^{th}$ instars of Aedes aegypti, but not against $4^{th}$ instars of Culex quinquefasciatus.

• Journal of Microbiology and Biotechnology
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• v.23 no.8
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• pp.1107-1115
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• 2013

The Cyt1Aa protein of Bacillus thuringiensis susbp. israelensis elaborates demonstrable toxicity to mosquito larvae, but more importantly, it enhances the larvicidal activity of this species Cry proteins (Cry11Aa, Cry4Aa, and Cry4Ba) and delays the phenotypic expression of resistance to these that has evolved in Culex quinquefasciatus. It is also known that Cyt1Aa, which is highly lipophilic, synergizes Cry11Aa by functioning as a surrogate membrane-bound receptor for the latter protein. Little is known, however, about whether Cyt1Aa can interact similarly with other Cry proteins not primarily mosquitocidal; for example, Cry2Aa, which is active against lepidopteran larvae, but essentially inactive or has very low toxicity to mosquito larvae. Here we demonstrate by ligand binding and enzyme-linked immunosorbent assays that Cyt1Aa and Cry2Aa form intermolecular complexes in vitro, and in addition show that Cyt1Aa facilitates binding of Cry2Aa throughout the midgut of C. quinquefasciatus larvae. As Cry2Aa and Cry11Aa share structural similarity in domain II, the interaction between Cyt1Aa and Cry2Aa could be a result of a similar mechanism previously proposed for Cry11Aa and Cyt1Aa. Finally, despite the observed interaction between Cry2Aa and Cyt1Aa, only a 2-fold enhancement in toxicity resulted against C. quinquefasciatus. Regardless, our results suggest that Cry2Aa could be a useful component of mosquitocidal endotoxin complements being developed for recombinant strains of B. thuringiensis subsp. israelensis and B. sphaericus aimed at improving the efficacy of commercial products and avoiding resistance.

• International Journal of Industrial Entomology and Biomaterials
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• v.36 no.2
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• pp.31-41
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• 2018

The dung beetle, Copris tripartitus (Coleoptera: Scarabaeidae), is an endangered insect in South Korea. Previously, partial mitochondrial COI and CytB gene sequences have been used to infer genetic diversity and gene flow of this species in South Korea. In this study, we additionally collected C. tripartitus (n = 35) from one previous locality and two new localities, sequenced COI and CytB genes, and combined these with preexisting data for population genetic analysis. Sequence divergence of current samples showed slightly lower values [4.86% (32 bp) for COI and 4.16% (18 bp) for CytB] than that in the previous study. Nucleotide diversity (${\pi}$) ranged from 0.005336 (Gulupdo) to 0.020756 (Seogwi-dong) in COI and 0.009060 (Aewol-eup) to 0.017464 (Seogwi-dong) in CytB. Seogwi-dong samples that showed the highest ${\pi}$ in the previous study also showed the highest ${\pi}$ in this study for both gene sequences. The newly investigated Gulupdo samples had the lowest haplotype diversity for both gene sequences. They also had the lowest ${\pi}$ for COI and the second lowest ${\pi}$ for CytB. On the other hand, the newly added Haean-dong sample had relatively higher diversity estimates. Gene flow among populations was high, although significant difference was only detected between Gulupdo and Anmado or between Gulupdo and Seogwi-dong for COI sequences (P < 0.05). Considering the high genetic diversity and gene flow in C. tripartitus populations, one major issue regarding conservation seems not to be recovery of genetic diversity.

• Journal of Sericultural and Entomological Science
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• v.37 no.2
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• pp.176-180
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• 1995

To characterize expression and formation of three type crystal proteins in transformant, Bacillus thruingiensis PT0529 was analysed by transmission electron microscope and SDS-PAGE according to growth. The results showed that the introduced crystal protein genes, rcyIVD and cytA, were well expressed at earlier stage than resident crystal proteins were also expressed with their own morphology. However, resident crystal protein of B. thuringiensis PT0529 was smaller than that of wild type B. thuringiensis NT0423, suggesting that resident crystal protein production was interfered with introduced two type crystal protein genes.

• Journal of Haehwa Medicine
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• v.19 no.2
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• pp.119-137
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• 2011

In order to improve efficacy of oriental medicines and to study the application of fermented oriental medicine in clinicals, the efficacy of CYT and CYTBH on atopic dermatitis were evaluated. The results and conclusions are as follows. CYT and CYTBH significantly improved the atopic dermatitis symptoms in NC/Nga mice by naked eye evaluation and significantly decreased clinical index in both groups. CYT and CYTBH both decreased the cell numbers of CD3+, CD11b+Gr-1+ cells in dorsal skin. Of the cells, CYT significantly decreased CD11b+Gr-1+ cells whereas CYTBH significantly decreased all immune cells. CYT and CYTBH both decreased the production rate of IL-4 and IFN-${\gamma}$ activated by CD3/CD28. In the case of CYTBH, significant decrease in all cases was observed. CYT and CYTBH decreased the production rate of IL-5, IL-13 and IL-17 in serum. Significant decrease of IL-5 in the case of CYT and IL-5 and IL-13 in the case of CYTBH were observed. CYT and CYTBH significantly decreased transcription of IL-5 mRNA and IL-13 mRNA in skin. Significant decrease in IgG1 and IgE immunoglobulins in serum were oberved in both groups. Significant decrease was only observed in the case of CYTBH. Both CYT and CYTBH significantly decreased the secretion of histamine. Both CYT and CYTBH suppressed erythema, hemorrhage, edema, excoriation, erosion of skin tissues of NC/Nga mice resulting in the decrease of thickness of epidermis. Significant decrease of infiltration of obese cells was also observed. The results above indicated that both CYT and CYTBH had significant efficacy in the treatment of atopic dermatitis through immune modulation. Animal studies showed that CYTBH had superior activity than that of CYT suggesting further and continuous studies on the changes in ingredients or absorption improvement by fermentation should follow.

• Fisheries and Aquatic Sciences
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• v.12 no.3
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• pp.171-178
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• 2009

The Perciformes include approximately 40% of all bony fishes and are the largest order of vertebrates. This order includes some of the most economically relevant marine fishes, particularly the red seabream, black seabream and rock bream. A 409 bp fragment of the cytochrome b (cyt b) gene and 403 bp and 518 bp fragments of ribosomal RNA (12S and 16S rRNA, respectively) were sequenced from five populations of natural and cultured red seabreams, natural black seabream, and natural and cultured rock breams. The mitochondrial DNA sequences were utilized for the genetic identification and population structural analyses of these three species. Phylogenetic relationships of intra- and inter-species were elucidated using three types of molecular genetic markers from three species of the order Perciformes in Korea. We noted no significant differences in the intra-specific variation of the cyt b and rRNA genes in each population however, inter-specific divergences were greater than intra-specific variation. Inter-specific variation was induced more by transition than transversion type in the cyt b and rRNA genes. The cyt b gene and rRNA genes make it possible to determine the inter-species divergence. The rRNA genes have more conserved sequences than the cyt b gene. Therefore, these genes are expected to prove useful among species belonging to the different genera or families.

• Molecules and Cells
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• v.25 no.4
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• pp.510-522
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• 2008

To construct the molecular systematics of the genus Megoura (Hemiptera: Aphididae), DNA based-identification was performed using four mitochondrial and three nuclear DNA regions: partial cytochrome c oxidase I (COI), partial tRNA-leucine + cytochrome c oxidase II (tRNA/COII), cytochrome b (CytB), partial 12S rRNA + tRNA-valine + 16S rRNA (12S/16S), elongation factor-1 alpha ($EF1{\alpha}$), and the internal transcribed spacers 1 and 2 (ITS1, ITS2). Pairwise sequence divergences between taxa were compared, and phylogenetic analyses were performed based on each DNA region separately, and the combined datasets. COI, CytB, $EF1{\alpha}$, ITS1, and ITS2 were relatively effective in determining species and resolving their relationships. By contrast, the sequences of tRNA/COII and 12S/16S were not able to separate the closely related species. CytB and $EF1{\alpha}$ gave better resolution with higher average sequence divergences (4.7% for CytB, 5.2% for $EF1{\alpha}$). The sequence divergence of COI (3.0%) was moderate, and those of the two ITS regions (1.8% for ITS1, 2.0% for ITS2) were very low. Phylogenetic trees were constructed by minimum evolution, maximum parsimony, maximum likelihood, and Bayesian phylogenetic analyses. The results indicated that the phylogenetic relationships between Megoura species were associated with their host preferences. Megoura brevipilosa and M. lespedezae living on Lespedeza were closely related, and M. nigra, monophagous on Vicia venosa, was rather different from M. crassicauda, M. litoralis, and M. viciae, which are oligophagous on Lathyrus and Vicia. The three populations of M. crassicauda formed a clade separated from M. litoralis and M. viciae. Nevertheless M. litoralis and M. viciae, which are morphologically similar, were not separated due to negligible sequence divergence. We discuss the phylogenetic relationships of the Megoura, and the usefulness of the seven DNA regions for determining the species level phylogeny of aphids.