• Animal cells and systems
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• v.13 no.3
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• pp.305-314
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• 2009

The control region of mitochondrial DNA (13516-14619) is located between srRNA and $tRNA^{lle}$ gene in swimming crab, Portunus trituberculatus. The present study was investigated the genetic polymorph isms of the control region in samples of P. trituberculatus collected at coastal waters of the Yellow Sea in Korea. A total of 300 substitution and indel polymorphic sites were identified. In addition to SNPs and indel variation, a hypervariable microsatellite motif was also identified at position from 14358 to 14391, which exhibited 10 alleles including 53 different suballeles. When the hypervariable microsatellite motif was removed from the alignment, 95 haplotypes were identified (93 unique haplotypes). The nucleotide and haplotype diversities were ranged from 0.024 to 0.028 and from 0.952 to 1.000, respectively. The statistically significant evidence for geographical structure was not detected from the analyses of neighbor-joining tree and minimum-spanning network, neither. This result suggest that population of P. trituberculatus are capable of extensive gene flow among populations. We believed that the polymorph isms of the control region will be used for informative markers to study phylogenetic relationships of P. trituberculatus.

• The Plant Pathology Journal
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• v.22 no.4
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• pp.369-374
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• 2006

To characterize benzimidazole-resistant and -sensitive Monilinia fructicola populations, the fungal isolates were obtained from peach plants showing brown rot and bloosom blight. Benzimidazole-sensitive isolates did not grow on potato dextrose agar(PDA) amended with $\geq1.0{\mu}g$ active ingredient(a.i.)/ml of the fungicides. However, benzimidazole-resistant isolates grew on PDA regardless of the tested concentrations of fungicides. Benzimidazole-resistant isolates did not grow on diethofencarb-PDA, but sensitive isolates grew on the same PDA. In the nucleotide sequences of $\beta$-tubulin gene, only codon 198(GAG: glutamic acid), a target site for benzimidazole, was replaced with GCG(alanine) in all of the resistant isolates, and this substitution seems to play an important role in the development of resistance. Other interesting codons such as 165(GCT), 200(TTC), and 241(GCT) were not changed among the isolates. Benzimidazole-resistant and -sensitive isolates were clustered clearly in random amplified polymerphic DNA analyses and the results revealed that low levels of genetic diversity between benzimidazole-sensitive and -resistant isolates of M. fructicola in the investigated regions.

• Korean Journal of Microbiology
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• v.24 no.4
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• pp.400-405
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• 1986

We have constructed a phylogenetic tree for eleven species by comparing their tRNA sequences. The tree suggests that prokaryotes diverged very early before the emergence of animals. The fact that H. volcano, an archaebacterium, clusters with eukaryotes implied that eukaryotes did not diverge directly from thier common ancestor with eubacteria. The branching order of phage $T_{4}$ and phage $T_{5}$ indicates that they have diverged separately from thier hosts and they might have evolved independently. A correlation between nucleotide substitution in tRNAs and paleontological record was observed. We verified that our phylogenetic tree fits very well with traditional ones very well by imposing the molecular clock on the tree.

• Toxicological Research
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• v.20 no.3
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• pp.187-193
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• 2004

This study aimed to determine the association of maternal oxidative stress and adverse pregnancy outcome with serum vitamin C concentration and a myeloperoxidase (MPO) genetic polymorphism during pregnancy. We investigated 450 pregnant women who visited Ewha Womans University Hospital for prenatal care during gestational weeks 24～28. During the second trimester, we measured serum vitamin C levels and urinary 8-hydroxyde-2'-deoxyguanosine (8-OHdG) and malondialdehyde (MDA) as an oxidative stress biomarker. We determined the presence of a maternal MPO polymorphism (G-to-A substitution at nucleotide 463) using a PCR-RFLP assay. We compared the level of oxidative stress and birth weight with the vitamin C concentration and the presence of the MPO polymorphism. The mean level of maternal oxidative stress tended to be higher and the birth weight lower for MPO type A/A than for types A/G and G/G. Vitamin C levels above the 75 percentiles were associated with reduced concentrations of urinary MDA and 8-OHdG but increased birth weight. Our data demonstrate that oxidative stress and neonatal birth weight are associated with the MPO genetic polymorphism, with the association modified by the maternal vita-min C levels.

• Molecules and Cells
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• v.23 no.2
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• pp.252-257
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• 2007

Escherichia coli HslVU is an ATP-dependent protease consisting of two heat shock proteins, the HslU ATPase and HslV peptidase. In the reconstituted enzyme, HslU stimulates the proteolytic activity of HslV by one to two orders of magnitude, while HslV increases the rate of ATP hydrolysis by HslU several-fold. Here we show that HslV alone can efficiently degrade certain unfolded proteins, such as unfolded lactalbumin and lysozyme prepared by complete reduction of disulfide bonds, but not their native forms. Furthermore, HslV alone cleaved a lactalbumin fragment sandwiched by two thioredoxin molecules, indicating that it can hydrolyze the internal peptide bonds of lactalbumin. Surprisingly, ATP inhibited the degradation of unfolded proteins by HslV. This inhibitory effect of ATP was markedly diminished by substitution of the Arg86 residue located in the apical pore of HslV with Gly, suggesting that interaction of ATP with the Arg residue blocks access of unfolded proteins to the proteolytic chamber of HslV. These results suggest that uncomplexed HslV is inactive under normal conditions, but may can degrade unfolded proteins when the ATP level is low, as it is during carbon starvation.

• Journal of Microbiology and Biotechnology
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• v.28 no.6
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• pp.1030-1036
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• 2018

Bacillus strains produce various types of antibiotics, and random mutagenesis has traditionally been used to overproduce these natural metabolites. However, this method leads to the accumulation of unwanted mutations in the genome. Here, we rationally designed a single nucleotide substitution in the degU gene to generate a B. subtilis strain displaying increased plipastatin production in a foreign DNA-free manner. The mutant strain (BS1028u) showed improved antifungal activity against Pythium ultimum. Notably, pps operon deletion in BS1028u resulted in complete loss of antifungal activity, suggesting that the antifungal activity strongly depends on the expression of the pps operon. Quantitative real-time PCR and lacZ assays showed that the point mutation resulted in 2-fold increased pps operon expression, which caused the increase in antifungal activity. Likewise, commercial Bacillus strains can be improved to display higher antifungal activity by rationally designed simple modifications of their genome, rendering them more efficient biocontrol agents.

• Parasites, Hosts and Diseases
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• v.45 no.4
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• pp.301-306
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• 2007

We attempted to identify parasite DNA in the biliary stones of humans via PCR and DNA sequencing. Genomic DNA was isolated from each of 15 common bile duct (CBD) stones and 5 gallbladder (GB) stones. The patients who had the CBD stones suffered from cholangitis, and the patients with GB stones showed acute cholecystitis, respectively. The 28S and 18S rDNA genes were amplified successfully from 3 and/or 1 common bile duct stone samples, and then cloned and sequenced. The 28S and 18S rDNA sequences were highly conserved among isolates. Identity of the obtained 28S D1 rDNA with that of Clonorchis sinensis was higher than 97.6%, and identity of the 18S rDNA with that of other Ascarididae was 97.9%. Almost no intra-specific variations were detected in the 28S and 18S rDNA with the exception of a few nucleotide variations, i.e., substitution and deletion. These findings suggest that C. sinensis and Ascaris lumbricoides may be related with the biliary stoneformation and development.

• Tuberculosis and Respiratory Diseases
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• v.62 no.5
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• pp.406-416
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• 2007

Background: Single nucleotide polymorphisms (SNPs), which consist of a substitution of a single nucleotide pair, are the most abundant form of genetic variations occurring with a frequency of approximately 1 per 1000 base pairs. SNPs by themselves do not cause disease but can predispose humans to disease, modify the extent or severity of the disease or influence the drug response and treatment efficacy. Single nucleotide polymorphisms (SNPs), particularly those within the regulatory regions of the genes often influence the expression levels and can modify the disease. Studies examining the associations between SNP and the disease outcome have provided valuable insight into the disease etiology and potential therapeutic intervention. Traditionally, the genotyping of SNPs has been carried out using polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP), which is a low throughput technique not amenable for use in large-scale SNP studies. Recently, TaqMan real-time PCR chemistry was adapted for use in allelic discrimination assays. This study validated the accuracy and utility of real-time PCR technology for SNPs genotyping Methods: The SNPs in promoter sequence (-37 and -524) of lung cancer suppressor gene, RRM1 (ribonucleotide reductase M1 subunit) with the genomic DNA samples of 89 subjects were genotyped using both real-time PCR and PCR-RFLP. Results: The discordance rates were 2.2% (2 mismatches) in -37 and 16.3% (15 mismatches) in -524. Auto-direct sequencing of all the mismatched samples(17 cases) were in accord with the genotypes read by real-time PCR. In addition, 138 genomic DNAs were genotyped using real-time PCR in a duplicate manner (two separated assays). Ninety-eight percent of the samples showed concordance between the two assays. Conclusion: Real-time PCR allelic discrimination assays are amenable to high-throughput genotyping and overcome many of the problematic features associated with PCR-RFLP.

• Microbiology and Biotechnology Letters
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• v.39 no.1
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• pp.93-96
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• 2011

It has been shown that a nucleotide substitution at position 770 in Escherichia coli 16S rRNA, which is implicated in forming the evolutionary conserved B2c intersubunit bridge, has a detrimental effect on ribosome function. In order to isolate second-site revertants that complement ribosomes containing C770G, we performed a random mutagenesis of the 16S rRNA gene and selected clones that could produce more CAT protein translated by specialized ribosome. One of the clones contained two nucleotide substitutions at positions 569 and 904 (C569G and U904C) and these mutations partially complemented the loss of protein-synthesis ability caused by C770G. Further studies using the isolated revertant will provide information about which part of 16S rRNA is interacting with C770 and the consequence of the structure formed by these interactions in the process of protein synthesis.

• Korean Journal of Medicinal Crop Science
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• v.14 no.3
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• pp.178-182
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• 2006

This study was conducted to investigate the variation in sequence, the base composition and the sequence similarity of 18S rDNA (18S ribosomal RNA coding region) in the 10 Polygonatum spp. collections. The entire 18S rDNA region of 10 Polygonatum spp. collections ranged from 913 bp to 914 bp. There were 8 variable sites in the entire 18S region, and they were attributable to nucleotide substitution and deletion. $T{\rightarrow}C$ transition happened in 4 sites, and $A{\rightarrow}G$ transition happened in 1 site. $C{\rightarrow}A$ transversion happened in 1 site, and deletion happened in 2 sites. Transition rates were five times that of transversion. Base compositions of 18S rDNA were $23.09{\sim}23.33%$ in adenine, $23.33{\sim}23.52%$ in guanine, $25.60{\sim}25.85%$ in thymine and $27.38{\sim}27.79%$ in cytosine. The A + T content of 18S rDNA of 10 Polygonatum spp. collections averages 48.99%, ranging from 48.80% to 49.18%, and the G + C content averages 51.01%, ranging from 50.82% to 51.20%. Pairwise sequence comparisons indicated that 18S rDNA sequence similarity ranged from 99.7% to 100%.