• Korean Journal of Microbiology
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• v.54 no.4
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• pp.420-427
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• 2018

The orthogonal pair of Saccharomyces cerevisiae tyrosyl-tRNA synthetase (Sc YRS) and a variant of E. coli initiator tRNA, fMam $tRNA_{CUA}$ which recognizes the amber stop codon is an effective tool for site-specific incorporation of unnatural amino acids into the protein in E. coli. To evolve the amber suppression activity of the orthogonal pair, we generated a mutant library of Sc YRS by randomizing two amino acids at 320 and 321 which involve recognition of the first base of anticodon in fMam $tRNA_{CUA}$. Two positive clones are selected from the library screening with chloramphenicol resistance mediated by amber suppression. They showed growth resistance against high concentration of chloramphenicol and their $IC_{50}$ values were approximately 1.7~2.3 fold higher than the wild type YRS. In vivo amber suppression assay reveals that mutant YRS-3 (mYRS-3) clone containing amino acid substitutions of P320A and D321A showed 6.5-fold higher activity of amber suppression compared with the wild type. In addition, in vitro aminoacylation kinetics of mYRS-3 also showed approximately 7-fold higher activity than the wild type, and the enhancement was mainly due to the increase of tRNA binding affinity. These results demonstrate that optimization of anticodon recognition by engineered aminoacyl tRNA synthetase improves the efficiency of unnatural amino acid incorporation in response to nonsense codon.

• Journal of Microbiology
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• v.45 no.6
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• pp.558-565
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• 2007

Using microarray analysis, we determined those Salmonella genes induced at the entry of stationary phase, and subsequently discovered that uncharacterized yliH was induced most dramatically. We set out to establish the molecular mechanism underlying the stationary phase induction of yliH under the standard culture condition, LB with vigorous aeration, by analyzing its promoter activity in various mutant backgrounds, lacking stationary phase ${\sigma}$, $RpoS^-$, or stringent signal molecules ppGpp, ${\Delta}relA$ ${\Delta}spoT$. It was found that the stationary phase induction of yliHp was partially dependent on rpoS but entirely dependent on ppGpp. DNA sequence analysis revealed that the Salmonella yliH gene is composed of 381 base-pair nucleotides, with overall amino acid sequence revealing 76.38% amino acid identity and 88.98% similarity with Escherichia coli yliH, although no motif from data base was noted for its possible role. Recently however, it has been reported that yliH in E. coli was implicated in biofilm formation and motility by repressing these activities (Domka et al., 2006). We have constructed a mutant Salmonella deleting yliH gene by allele replacement and examined its phenotype, and found that the yliH in Salmonella more or less affects motility and adherence by enhancing these activities. The effect on biofilm formation in Salmonella was uncertain. Moreover, addition of cloned yliH of E. coli into Salmonella did not reduce motility or adherence. Taken together, it appears that the pathways implicating yliH for biofilm formation and motility in E. coli and in Salmonella are somewhat different.

• Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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• 2008.05a
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• pp.616-619
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• 2008

In this paper, we design and implement the authentication system for home network service and applied it to actual sensor nodes. We achieved authentication key, encryption and decryption applied RC5 encryption algorithm of SNEP. In addition, we used pair-wise key pre-distribution for prevention of authentication sniffing in wireless sensor network. The experiment environment consists of a base station receiving data and sensor nodes sending data. Each sensor nodes sends both the data and encrypted authentication key to the base station. As a simulation environment, we assumed some what-if scenarios of security menaces in home network service. And we slightly altered the TOS_Msg construction of TinyOS. The experiences had shown that the malfunction doesn't happen in communication among other groups. And we confirmed in tests that the system is secure when a sensor having malicious propose is added.

• Journal of Life Science
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• v.15 no.5 s.72
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• pp.734-738
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• 2005

A gene coding for xylanase from alkali-tolerant Bacillus alcalophilus AX2000 was cloned into Escherichia coli $DH5\alpha$ using pUC19. Among 2,000 transformants, one transformant showed clear zone on the detection agar plate containing oat-spells xylan. Its recombinant plasmid, named pXTY99, was found to carry 7.0 kb insert DNA fragment. When the nucleotide sequence of the cloned xylanase gene (xynT) was determined, xynT gene was found to consist of 1,020 base-pair open reading frame coding for a poly-peptide of 340 amino acids with a deduced molecular weight of 40 kDa. The coding sequence was preceded by a putative ribosome binding site, and the transcription initiation signals. The deduced amino acid sequence of xylanase is similar to those of the xylanases from Bacillus sp. Nl37 and B. stearothermophilus 21 with $61\%$ and $59\%$ identical residues, respectively.

• The Plant Pathology Journal
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• v.18 no.2
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• pp.98-101
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• 2002

Using phytoplasma universal primer pair Pl and P7, a fragment of about 1.8 kb nucleotide sequences of 16S rRNA gene and 16S-23S rRNA intergenic spacer region, and a portion of 23S rRNA gene of jujube witches'broom (JWB) and mulberry dwarf(MD) phytoplasmas were determined. The nucleotide sequences of JWB and MD were 1,850 bp and 1,831 bp long, respectively. The JWB phytoplasma sequence was aligned with the homologous sequence of MD phytoplasma. Twenty-eight base insertions and nine base deletions were found in the JWB phytoplasma sequence compared with that of MD phytoplasma. The similarity of the aligned sequences of JWB and MD was 84.8%. The near-complete 16S rRNA gene DNA sequences of JWB and MD were 1,529 bp and 1,530 bp in length, respectively, and revealed 89.0% homology. The 16S-23S rRNA intergenic spacer region DNA sequences were 263 bp and 243 bp in lengths respectively, while homology was only 70% and the conserved tRNA-lle gene of JWB and MD was located into the intergenic space region between 16S-23S rRNA gene. The nucleotide sequences were 77 bp long in both JWB and MD, and showed 97.4% sequence homology. Based on the phylogenetic analysis of the two phytoplasmas, the JWB phytoplasma belongs to the Elm yellow phytoplasma group (16S rV), whereas, the MD phytoplasma belongs to the Aster yellow group (16S rI).

• Molecules and Cells
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• v.27 no.3
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• pp.365-381
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• 2009

The chloroplast DNA sequences of Megaleranthis saniculifolia, an endemic and monotypic endangered plant species, were completed in this study (GenBank FJ597983). The genome is 159,924 bp in length. It harbors a pair of IR regions consisting of 26,608 bp each. The lengths of the LSC and SSC regions are 88,326 bp and 18,382 bp, respectively. The structural organizations, gene and intron contents, gene orders, AT contents, codon usages, and transcription units of the Megaleranthis chloroplast genome are similar to those of typical land plant cp DNAs. However, the detailed features of Megaleranthis chloroplast genomes are substantially different from that of Ranunculus, which belongs to the same family, the Ranunculaceae. First, the Megaleranthis cp DNA was 4,797 bp longer than that of Ranunculus due to an expanded IR region into the SSC region and duplicated sequence elements in several spacer regions of the Megaleranthis cp genome. Second, the chloroplast genomes of Megaleranthis and Ranunculus evidence 5.6% sequence divergence in the coding regions, 8.9% sequence divergence in the intron regions, and 18.7% sequence divergence in the intergenic spacer regions, respectively. In both the coding and noncoding regions, average nucleotide substitution rates differed markedly, depending on the genome position. Our data strongly implicate the positional effects of the evolutionary modes of chloroplast genes. The genes evidencing higher levels of base substitutions also have higher incidences of indel mutations and low Ka/Ks ratios. A total of 54 simple sequence repeat loci were identified from the Megaleranthis cp genome. The existence of rich cp SSR loci in the Megaleranthis cp genome provides a rare opportunity to study the population genetic structures of this endangered species. Our phylogenetic trees based on the two independent markers, the nuclear ITS and chloroplast MatK sequences, strongly support the inclusion of the Megaleranthis to the Trollius. Therefore, our molecular trees support Ohwi's original treatment of Megaleranthis saniculifolia to Trollius chosenensis Ohwi.

• Fisheries and Aquatic Sciences
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• v.18 no.1
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• pp.99-107
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• 2015

A natural hybrid of a probable intergeneric mating between the striped shiner Pungtungia herzi and the stone morocco Pseudorasbora parva (Cypriniformes: Cyprinidae) was captured in the Geumho River, a tributary of the Nakdong River basin in Korea. Morphological characters and DNA sequences were analyzed to verify its hybrid state and identify the parentage of its parent species. The hybrid exhibited a phenotypic intermediacy between the two parent species in the number of vertebrae and the mouth shape. Out of 1,488 base pair (bp) positions of the nuclear recombination activating gene 1 gene (rag1), which has a biparental mode of inheritance, 41-bp substitutions were detected between the two parent species, whereas an electropherogram of the hybrid displayed polymorphic double peaks at all of the base positions, along with one additional one, strongly indicating its hybrid state. Meanwhile, sequence comparison of the mitochondrial cytochrome b gene (mt-cyb) (1,140 bp), which has a maternal mode of inheritance, showed only 5-22-bp differences (97.6-99.5% identities) between the hybrid and Pu. herzi, but as many as 158-168-bp differences (85.2-86.1% identities) between the hybrid and Ps. parva, clearly indicating Pu. herzi as the maternal species. Thus, combined analyses of independent data sets (i.e., morphology and nuclear and mitochondrial DNA sequences) offered convincing evidence for the hybrid state of a naturally occurring hybrid resulting from intergeneric mating between a female Pu. herzi and a male Ps. parva.

• Journal of Ecology and Environment
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• v.36 no.3
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• pp.159-166
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• 2013

Oryzias latipes and Oryzias sinensis are indigenous species found in Japan, China, and other East Asian countries, including Korea. Based on morphological differences, the species have been classified distinctly. However, the range of morphological characters such as the number of gill rakers, vertebrae, and spots on the lateral body overlaps and is too vague for clear identification, so their classification based on their morphological characteristics remains uncertain. In this study, the mitochondrial cytochrome oxidase subunit I (COI) gene, which is used for DNA barcoding, was applied to clarify interspecific variation of O. latipes and O. sinensis. Intraspecific genetic diversity was calculated to identify correlations with geographic distributions. We studied two species collected from 55 locations in Korea. All individuals carried a 679-base pair gene without deletion or insertion. Between species, 525 base pairs of the gene were shared. The Kimura two parameter (K2P) distance of O. latipes and O. sinensis was 0.41% and 1.39%, respectively. Mean divergence within genera was 23.5%. Therefore, the species were clearly different. The distance between O. latipes and O. sinensis was 14.0%, which is the closest within genera. Interestingly O. latipes from the Japanese and Korean group represented 16.5% distant. These results were derived from geohistorical and anthropogenic environmental factors. The O. latipes haplotypes were joined in only one group, but O. sinensis was divided into two groups, one is found in the Han River and upper Geum River watershed; the other is found in the remaining South Korean watersheds. Further studies will address the causes for geographic speciation of O. sinensis haplotypes.

• Journal of The Korean Society of Inherited Metabolic disease
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• v.12 no.1
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• pp.54-57
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• 2012

A 3-year-old boy was transferred to emergency room (ER) with lethargy and abdominal pain. Physical examination revealed drowsy mental status but neurologically intact. Basic evaluation in ER shows hypoglycemia (43 mg/dL), hyperglycerolemia, ketonemia and ketonuria. Initial urine organic acid was performed and the result showed severe hyperglyceroluria. Under suspicion of isolated GKD, GKD gene was obtained from his DNA from white blood cell in peripheral blood and sequencing was performed. Isolated glycerol kinase deficiency (GKD) is an X-linked inborn error of metabolism that is either symptomatic or asymptomatic. GKD is due to deletions of, or mutations within, the GK gene, and there is no genotype-phenotype correlation. Gene study that we performed showed normal at a well-known mutation site, but found 4-base insertion at 79 base pair away from the beginning of exon 11.

• Animal cells and systems
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• v.14 no.2
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• pp.137-145
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• 2010

Four specimens of unknown Coilia sp. were collected for the first time from the Yellow Sea in 2008 and compared with Coilia mystus and Coilia nasus. Coilia sp. showed similar morphology to C. mystus and C. nasus, but differed in that its tail was considerably shorter. We conducted an analysis of the morphological and genetic characteristics in an effort to clarify the taxonomic position of Coilia sp. In counts and measurements, Coilia sp. were well distinguished from C. nasus by the number of scutes (42-44 in Coilia sp. vs. 40-45 in C. mystus vs. 45-55 in C. nasus), ratio of dorsal base length to head length (43.4-47.6 vs. 37.9-47.6 vs. 33.0-41.0), and eye length to head length (19.2-20.8 vs. 17.0-22.4 vs. 13.8-18.2). In caudal skeleton of Coilia sp., urostyle, hypural and epural bones were not observed; instead of them, caudal fin rays were supported by the last vertebra, neural and haemal spines' extension. The molecular phylogenetic relationship was analyzed using 414 base-pair 12S rRNA mitochondrial DNA sequences. The Kimura-2-parameter distance between Coilia sp. and C. mystus was 0.3%, but was 1.3% between Coilia sp. and C. nasus. Both the neighbor-joining tree and maximum-likelihood tree showed that Coilia sp. are closely clustered with C. mystus. Therefore, our results suggest that the Coilia sp. may be a deformed fish of C. mystus.