• Fisheries and Aquatic Sciences
• /
• 제12권3호
• /
• pp.171-178
• /
• 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.

• BMB Reports
• /
• 제43권6호
• /
• pp.427-431
• /
• 2010

Cyt2Aa2 is a mosquito-larvicidal protein produced as a 29 kDa crystalline protoxin from Bacillus thuringiensis subsp. darmstadiensis. To become an active toxin, proteolytic processing is required to remove amino acids from its N- and C-termini. This study aims to investigate the functional role of amino acid residues on the N-terminal ${\beta}1$ and C-terminal ${\alpha}F$ of Cyt2Aa2 protoxin. Mutant protoxins were constructed, characterized and compared to the wild type Cyt2Aa2. Protein expression data and SDS-PAGE analysis revealed that substitution at leucine-33 (L33) of ${\beta}1$ has a critical effect on dimer formation and structural stability against proteases. In addition, amino acids N230 and I233-F237 around the C-terminus ${\alpha}F$ demonstrated a crucial role in protecting the protoxin from proteolytic digestion. These results suggested that ${\beta}1$ and ${\alpha}F$ on the Nand C-terminal ends of Cyt2Aa2 protoxin play an important role in the molecular interaction and in maintaining the structural stability of the protoxin.

• 대한한방내과학회지
• /
• 제20권1호
• /
• pp.99-110
• /
• 1999

Effects of Chiyangtang(CYT) on H. pylori-induced increase of interleukin 8 and interleukin 1 gene expression was studied in Kato Ⅲ cell line, a human stomach epithelial cell line. Treatment of H. pylori to the cell culture signifant!y increased IL-8 and IL-1 mRNA synthesis. When CYT was added along with H. pylori, the increase of IL-8 and IL-1 mRNA synthesis was blocked. Activation of transcription factor $NF-{\kappa}B$ and AP-1 which were known to important in IL-8 and IL-1 gene expression was also studied using chloramphenicol acetyltransferase(CAT) assay. Treatment of H. pylori increased activation of $NF-{\kappa}B$ and AP-l and CYT effectively protected the activation. Electrophoretic mobility shift assay suggested that CYT effectively inhibited DNA binding of $NF-{\kappa}B$ and AP-l to their cognate site. These results suggested that CYT could prevent stomach diseases through the down regulation of IL -8 and IL-l gene expression which might be mediated by the inhibition of $NF-{\kappa}B$ and AP-1 activities and their binding to DNA.

• Molecules and Cells
• /
• 제25권4호
• /
• pp.510-522
• /
• 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.

• Animal Systematics, Evolution and Diversity
• /
• 제29권2호
• /
• pp.136-143
• /
• 2013

The collared scops owl that occurs in Korea is a protected species but its exact specific status has been questioned. To resolve the species status, a molecular phylogenetic analysis was conducted using two fragments of mitochondrial DNA, cytochrome b (cyt b, 891 bp) and NADH dehydrogenase subunit 2 (ND2, 627 bp) genes. Phylogenetic trees of cyt b revealed that all Korean specimens formed a monophyletic group with Japanese scops owl Otus semitorques with very low sequence divergence (d=0.008). We obtained a similar ND2 tree as well (d=0.003); however, the genetic distance between Korean individuals and O. lempiji from GenBank (AJ004026-7, EU348987, and EU601036) was very high and sufficient enough to separate them as species (cyt b, d=0.118; ND2, d=0.113). We also found that Korean species showed high differentiation from O. bakkamoena (AJ004018-20 and EU601034; cyt b, d=0.106; ND2, d=0.113) and O. lettia (EU601109 and EU601033, cyt b, d=0.110; ND2, d=0.117) as well. Therefore, we suggest that the Korean collared scops owl should be designated as Otus semitorques.

• Applied Biological Chemistry
• /
• 제51권4호
• /
• pp.263-268
• /
• 2008

Seventy-one Bacillus cereus strains (12 references and 59 food isolates) were analyzed for the occurrence of five different enterotoxin genes (nheABC, hblCDA, entFM, cytK, and bceT) and one emetic toxin cereulide synthetase gene (ces) by PCR (polymerase chain reaction). PCR analysis revealed eight toxigenic patterns in all B. cereus strains tested; they all carried both entFM and nheABC. The presence of hblCDA, cytK, and bceT varied according to the enterotoxin-producing strains, among which hblCDA was the least frequently detected in the food-isolated strains. Only five B. cereus strains harbored ces, associated with the emetic type of food poisoning; however, these strains were devoid of hblCDA, cytK, and bceT.

• Bulletin of the Korean Chemical Society
• /
• 제17권3호
• /
• pp.279-284
• /
• 1996

Decay of the spin label attached to cytochrome c or to stearic acid has been measured by electron paramagnetic resonance (EPR) spectroscopy to monitor membrane oxidation induced by cytochrome c-membrane interaction. Binding of cytochrome c sequestered the acidic phospholipids and membrane oxidation was efficient in the order linoleic oleic>stearic acid for a fatty acid chain in the acidic phospholipids. The spin label on cyt c was destroyed at pH 7 whereas that on stearic acid embedded in the membrane was destroyed at pH 4, presumably due to different modes of cyt c-membrane interaction depending on pH. Interestingly, cyt c also interacts with phosphatidylethanolamine, an electrically neutral phospholipid, to cause rapid membrane oxidation. Both EPR and fluorescence measurements indicated that electrostatic interaction is at least partially responsible for the process.

• Applied Biological Chemistry
• /
• 제41권1호
• /
• pp.23-30
• /
• 1998

B. thuringiensis에서 분리된 cryIB, truncated cryIB$[cryIB({\alpha})]$, cryIA(b), 및 cytA 유전자를 E. coli-Bacillus의 shuttle vector인 pBES에 cloning하여 이 유전자의 동종 혹은 이종발현과 이들 형질전환된 균주에서 생성되는 살충성 결정 단백질의 형태와 특성을 조사하였다. E. coli와 Bacillus의 형질전환은 electroporation으로 이루어졌으며, 효과적인 형질전환을 위한 field strength와 resistance는 E. coli의 경우 11.0 kV/cm와 129 ohms였고, Bacillus의 경우에는 4.5 kV/cm와 48 ohms였다. E. coli나 Bacillus에서 형성되는 살충성 결정 단백질의 전자현미경적인 형태는 원래의 숙주에서 형성된 것과 동일하여 CryIB와 CryIA(b)는 bipyramid 형이고, CytA는 부정형이었으며, 크기는 E. coli에서 형성된 것이 Bacillus에서 형성된 것 보다 작았다. Alkaline pH에서의 살충성 결정 단백질의 용해도는 E. coli의 경우 pH의 증가에 따라 점진적으로 증가되었으나, Bacillus의 경우에는 pH 9 까지는 점진적으로 증가하다가 pH 9 이상에서는 큰 폭으로 증가하여 E. coli의 경우보다 2배 이상의 차이를 보였다.

• Asian-Australasian Journal of Animal Sciences
• /
• 제26권2호
• /
• pp.163-170
• /
• 2013

In order to analyze the genetic diversity and phylogenetic status of the Korean Chikso breed, we determined sequences of mtDNA cytochrome b (cyt b) gene and performed phylogenetic analysis using 239 individuals from 5 Chikso populations. Five non-synonymous mutations of a total of 15 polymorphic sites were identified among 239 cyt b coding sequences. Thirteen haplotypes were defined, and haplotype diversity was 0.4709 ranging from 0.2577 to 0.6114. Thirty-five haplotypes (C1-C35) were classified among 9 Asia and 3 European breeds. C2 was a major haplotype that contained 206 sequences (64.6%) from all breeds used. C3-C13 haplotypes were Chikso-specific haplotypes. C1 and C2 haplotypes contained 80.5% of cyt b sequences of Hanwoo, Yanbian, Zaosheng and JB breeds. In phylogenetic analyses, the Chikso breed was contained into B. taurus lineage and was genetically more closely related to two Chinese breeds than to Korean brown cattle, Hanwoo. These results suggest that Chikso and Hanwoo have a genetic difference based on the mtDNA cyt b gene as well as their coat color, sufficient for classification as a separate breed.

• The Korean Journal of Physiology and Pharmacology
• /
• 제14권6호
• /
• pp.407-412
• /
• 2010

3-Deazaadenosine (DZA), a potent inhibitor of S-adenosylhomocysteine hydrolase, was previously proposed to induce intrinsic apoptosis in human leukemic cells. In the present study, we analyzed the mechanism underlying the DZA-induced intrinsic apoptotic pathway. DZA activated typical caspase-dependent apoptosis in HL-60 cells, as demonstrated by an accumulation of hypo-diploidic cells, the processing of multiple procaspases and an inhibitory effect of z-VAD-Fmk on this cell death. During DZA-induced apoptosis, cytochrome c (cyt c) was released into the cytosol. This was neither prevented by z-VAD-Fmk and nor was it associated with the dissipation of mitochondrial membrane potential (${\Delta}{\Psi}_m$). Prior to the release of cyt c, BAX was translocated from the cytosol to mitochondria and underwent oligomerization. Finally, the overexpression of BCL-XL protected HL-60 cells from apoptosis by blocking both the cyt c release and BAX oligomerization. Collectively, these findings suggest that DZA may activate intrinsic apoptosis by stimulating BAX activation and thereby the release of cyt c.