• Asian-Australasian Journal of Animal Sciences
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• v.31 no.9
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• pp.1516-1524
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• 2018

Objective: Defensins are a large family of antimicrobial peptides and components of the innate immune system that invoke an immediate immune response against harmful pathogens. Defensins are classified into alpha-, beta-, and theta-defensins. Avian species only possess beta-defensins (AvBDs), and approximately 14 AvBDs (AvBD1-AvBD14) have been identified in chickens to date. Although substantial information is available on the conservation and phylogenetics, limited information is available on the expression and regulation of AvBD8 in chicken immune tissues and cells. Methods: We examined AvBD8 protein expression in immune tissues of White Leghorn chickens (WL) by immunohistochemistry and quantitative reverse transcription-polymerase chain reaction (RT-qPCR). In addition, we examined AvBD8 expression in chicken T-, B-, macrophage-, and fibroblast-cell lines and its regulation in these cells after lipopolysaccharide (LPS) treatment by immunocytochemistry and RT-qPCR. Results: Our results showed that chicken AvBD8 protein was strongly expressed in the WL intestine and in macrophages. AvBD8 gene expression was highly upregulated in macrophages treated with different LPS concentrations compared with that in T- and B-cell lines in a time-independent manner. Moreover, chicken AvBD8 strongly interacted with other AvBDs and with other antimicrobial peptides as determined by bioinformatics. Conclusion: Our study provides the expression and regulation of chicken AvBD8 protein in immune tissues and cells, which play crucial role in the innate immunity.

• BMB Reports
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• v.41 no.6
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• pp.466-471
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• 2008

Three isoforms of pig PDE4B were cloned and classified as two forms: PDE4B1 and PDE4B3, which contain UCR1 and UCR2; and PDE4B2, which contains only UCR2. The amino acid sequences of each isoform showed good conservation in human and rat. PDE4B2 is expressed in a wide range of tissues, but PDE4B1 and PDE4B3 are not. Using an informative SNP for the Iberian x Landrace intercross detected from intron 12, a linkage map was constructed. The location of PDE4B was estimated at 123.6 cM outside of the QTL-CI (124-128 cM) for IMF. However, the QTL-CI for IMF was reconfirmed with high significance, and its position was narrowed down to an interval of 4 cM (the region defined by markers PDE4B and SW1881). Using radiation hybrid mapping, LEPR, LEPROT, DNAJC6, AK3L1 and AK3L2 were selected as positional and/or functional candidates related to the QTL.

• Journal of Microbiology and Biotechnology
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• v.28 no.7
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• pp.1185-1198
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• 2018

The nucleosomal organization of chromatin using histone proteins is a fundamental and ubiquitous feature of eukaryotic nuclei, with the major exception of dinoflagellates. Although a number of recent genomic and transcriptomic analyses have detected numerous histone genes in dinoflagellates, little is known about their expression. Here in, we aimed to investigate the expression pattern of histone genes under nutritional stress, and an attempt was made to detect histone expression at the protein level in Alexandrium pacificum. The presence of histones at the mRNA level was confirmed in this study by the amplification, cloning, and sequencing of 10 different genes. Relative expression profiling of these genes under different growth conditions was determined with real-time PCR and revealed considerable levels of histone transcription in nutritionally stressed cells. We were unable to detect the expression of histones at the protein level even after immunodetection and analysis using mass spectrometry, although a histone-like protein was detected as a major nuclear component. A. pacificum expresses multiple variants of histone, and protein sequences revealed both conservation and divergence with respect to other eukaryotes. We concluded that A. pacificum maintained an active transcription of histone genes within the cell, and enhanced expression of histone genes in nutritional stress strongly suggest that histones have functional significance in dinoflagellates, although expression at the protein level was below our current detection limits, which suggests a limited role of histones in DNA packaging. Finally, the plausible regulation of histone expression at the gene and protein levels in A. pacificum is discussed.

• Fisheries and Aquatic Sciences
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• v.13 no.3
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• pp.224-230
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• 2010

Growth hormone (GH) is known as one of the main osmoregulators in euryhaline teleosts during seawater (SW) adaptation. Many of the physiological actions of GH are mediated through insulin-like growth factor-I (IGF-I), and the GH/IGF-I axis is associated with osmoregulation of fish during SW acclimation. However, little information is available on the response of fish IGF-II to hyperosmotic stress. Here we present the first cloned IGF-I and IGF-II cDNAs of marine medaka, Oryzias dancena, and an analysis of the molecular characteristics of the genes. The marine medaka IGF-I cDNA is 1,340 bp long with a 257-bp 5' untranslated region (UTR), a 528 bp 3' UTR, and a 555-bp open reading frame (ORF) encoding a propeptide of 184 amino acid (aa) residues. The full-length marine medaka IGF-II cDNA consists of a 639 bp ORF encoding 212 aa, a 109 bp 5' UTR, and a 416 bp 3' UTR. Homology comparison of the deduced aa sequences with other IGF-Is and IGF-IIs showed that these genes in marine medaka shared high structural homology with orthologs from other teleost as well as mammalian species, suggesting high conservation of IGFs throughout vertebrates. The IGF-I mRNA level increased following transfer of marine medaka from freshwater (FW) to SW, and the expression level was higher than that of the control group, which was maintained in FW. This significantly elevated IGF-I level was maintained throughout the experiment (14 days), suggesting that in marine medaka, IGF-I is deeply involved in the adaptation to abrupt salinity change. In contrast to IGF-I, the increased level of marine medaka IGF-II mRNA was only maintained for a short period, and quickly returned a level similar to that of the control group, suggesting that marine medaka IGF-II might be a gene that responds to acute stress or one that produces a supplemental protein to assist with the osmoregulatory function of IGF-I during an early phase of salinity change.

• Plant Biotechnology Reports
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• v.4 no.1
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• pp.15-21
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• 2010

The over-expression of Arabidopsis CAX1 and CAX2 causes transgenic tomato plants to reveal severe $Ca^{2+}$ deficiency-like symptoms such as tip-burn and/or blossom end rot, despite there being sufficient $Ca^{2+}$ in each plant part. To correct the symptoms and to moderately enhance the calcium level, a worldwide vegetable tomato was genetically engineered using a modified Arabidopsis cation/$H^+$ antiporter sCAX2A, a mutant form of Arabidopsis CAX2. Compared with the wild-type, the sCAX2A-expressing tomato plants demonstrated elevated $Ca^{2+}$ levels in the fruits with almost no changes in the levels of $Mn^{2+}$, $Cu^{2+}$, and $Fe^{2+}$. Moreover, expression of sCAX2A in tomato plants did not show any significant alterations in their morphological phenotypes. Unlike 35S::sCAX1 construct, sCAX2A antiporter gene driven by 35S promoter can be a valuable tool for enriching $Ca^{2+}$ contents in the tomato fruit without additional accumulation of the undesirable cations.

• Fisheries and Aquatic Sciences
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• v.11 no.2
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• pp.82-87
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• 2008

Marine fisheries are important natural resources and must be maintained, especially fish species that are important sources of food. Despite the increase in stocking programs to maintain fisheries with artificially raised fish, the genetic impact stocking has on the wild fry population has not been addressed. Genetic variation in rock bream, Oplegnathus fasciatus, within and between wild-caught parents and the $F_1$ generation produced by them in 1 day was assayed using nine highly variable micro satellite markers. The nine micro satellite loci used in this study displayed diverse polymorphisms, and in total, 98 different alleles were observed over all loci. Differences in genetic variability of the $F_1$ offspring compared to their wild-caught parents (brood stock) were observed in terms of allele frequency, gene diversity, and heterozygosity. Although the $F_1$ generation of rock bream was missing 16% of the micro satellite alleles, no significant reduction was found in mean heterozygosity of the $F_1$ population compared to the brood stock. Eight of nine loci showed significant Hardy-Weinberg equilibrium (HWE) deviations in the $F_1$ population, while the brood stock deviated from HWE at three micro satellite loci (KOF85, KOF360 and KOF374). These deviations showed mostly a deficit of heterozygotes. Our results provide evidence for genetic differences in the $F_1$ hatchery offspring compared to their wild-caught parents and reinforce the need for a series of consecutive egg collections to avoid the loss of genetic variability. This also further underscores the importance of monitoring genetic variability of hatchery populations for the conservation of natural rock bream resources.

• Plant Resources
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• v.6 no.2
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• pp.120-128
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• 2003

The objectives of this study, an analysis of the variation for leaf and fruit characteristics among the selected ten populations of Sorbus commixta Hedl. could be used for the conservation of gene resources and could provide information to superior trees selection. The results obtained from this study can be summarized as follows; Approximately, the Mt. Sungin population at Ulleung island showed larger values in overall characteristics and populations. On the other hand, Mt. Halla population at Jeju island showed the smaller values of the overall characteristics and populations. ANOV A tests showed that there were statistically significant differences in all leaf characteristics among the populations as well as individual trees within populations. But, for fruit characteristics, differences were statistically significant only among the populations. Cluster analysis using single linkage method based on leaf and fruit characteristics showed that ten selected populations of S. commixta in Korea could be clustered into three groups. Group I is Mt. Sungin at Ulleung island, Group II is Mt. Halla at Jeju island, and Group III comprises Osan, Mt. Kaji, Mt. Duckyoo, Mt. Balwang, Mt. Sobaek, Mt. O-dae, Mt. Jiri, and Mt. Taebaek. The selection level based on major agronomic traits, which are the Number of Fruit per Fruiting Lateral(NFL) over 50, and Fruit Length(FL) and Width(FW) over 10 mm, and Weight of 100 Fruit(WFI00) over 66 g, was applied on 100 sample trees, and five trees were selected. The selection effects from selected trees in NFL, FL, FW, and WF100 were evaluated as 132%, 151 %, 142%, and 264% compared to the mean of those 100 sample trees, respectively. Especially, Ulleung 2 showed excellent values that NFL and WFI00 were 95, and 69 g, respectively, suggesting a promising new cultivar for larger fruit and high productivity.

• Journal of Microbiology and Biotechnology
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• v.24 no.11
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• pp.1551-1558
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• 2014

The esterase gene Est8 from the thermophilic bacterium Bacillus sp. K91 was cloned and expressed in Escherichia coli. The monomeric enzyme exhibited a theoretical molecular mass of 24.5 kDa and an optimal activity around $50^{\circ}C$ at pH 9.0. A model of Est8 was constructed using a hypothetical YxiM precursor structure (2O14_A) from Bacillus subtilis as template. The structure showed an ${\alpha}/{\beta}$-hydrolase fold and indicated the presence of a typical catalytic triad consisting of Ser-11, Asp-182, and His-185, which were investigated by site-directed replacements coupled with kinetic characterization. Asp-182 and His-185 residues were more critical than the Ser-11 residue in the catalytic activity of Est8. A comparison of the amino acid sequence showed that Est8 could be grouped into the GDSL family and further classified as an SGNH hydrolase. Est8 is a new member of the SGNH hydrolase subfamily and may employ a different catalytic mechanism.

• Korean Journal of Plant Taxonomy
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• v.38 no.1
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• pp.51-61
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• 2008

Leaves are indeterminate organs and possess a lot of genes which is involved in establishing leaf polarities. These polarities are regulated relatively early during leaf development and defined relative to the factors intrinsic to the primordia and interactions with the shoot apical meristem (SAM). Recently, several genes that control the polarity of lateral organs have been identified. Our genetic study of deformed root and leaf1 (drl1) mutant, which produces narrow, filament‐like leaves and defective meristems, revealed that DRL1 is involved in the regulation of SAM activity and leaf polarity. The DRL1 gene was found to encode a novel protein showing homology to Elongator‐associate protein (EAP) of yeast KTI12. The amino acid sequence of DRL1 is universally conserved in prokaryotes and eukaryotes. DRL1 and the plant DRL1 homologs clearly formed a monophyletic clade, suggesting the evolutionary conservation of DRL1 homologs was maintained in the genomes of all land plants.

• Journal of Microbiology and Biotechnology
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• v.18 no.7
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• pp.1235-1244
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• 2008

Indole-3-acetic acid (IAA) is produced commonly by plants and many bacteria, however, little is known about the genetic basis involving the key enzymes of IAA biosynthetic pathways from Bacillus spp. IAA intermediates from the Gram-positive spore-forming bacterium Paenibacillus polymyxa E681 were investigated, which showed the existence of only an indole-3-pyruvic acid (IPA) pathway for IAA biosynthesis from the bacterium. Four open reading frames (ORFs) encoding indole-3-pyruvate decarboxylase-like proteins and putative indole-3-pyruvate decarboxylase (IPDC), a key enzyme in the IPA synthetic pathway, were found on the genome sequence database of P. polymyxa and cloned in Escherichia coli DH5$\alpha$. One of the ORFs, PP2_01257, was assigned as probable indole-3-pyruvate decarboxylase. The ORF consisted of 1,743 nucleotides encoding 581 amino acids with a deduced molecular mass of 63,380 Da. Alignment studies of the deduced amino acid sequence of the ORF with known IPDC sequences revealed conservation of several amino acids in PP2_01257, essential for substrate and cofactor binding. Recombinant protein, gene product of the ORF PP2_01257 from P. polymyxa E681, was expressed in E. coli BL21 (DE3) as a glutathione S-transferase (GST)-fusion protein and purified to homogeneity using affinity chromatography. The molecular mass of the purified enzyme showed about 63 kDa, corresponding closely to the expected molecular mass of IPDC. The indole-3-pyruvate decarboxylase activity of the recombinant protein, detected by HPLC, using IPA substrate in the enzyme reaction confirmed the identity and functionality of the enzyme IPDC from the E681 strain.