• The Plant Pathology Journal
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• v.25 no.4
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• pp.344-351
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• 2009

A biocontrol bacterium Bacillus licheniformis N1 grown in nutrient broth showed no chitinolytic activity, while its genome contains a gene which encodes a chitinase. The gene for chitinase from B. licheniformis N1 was amplified by PCR and the deduced amino acid sequence analysis revealed that the chitinase exhibited over 95% identity with chitinases from other B. licheniformis strains. Escherichia coli cells carrying the recombinant plasmid displayed chitinase activity as revealed by the formation of a clear zone on chitin containing media, indicating that the gene could be expressed in E. coli cells. Chitinase gene expression in B. licheniformis N1 was not detected by RT-PCR analysis. The protein was over-expressed in E. coli BL21 (DE3) as a glutathione S-transferase fusion protein. The protein could also be produced in B. subtilis 168 strain carrying the chitinase gene of N1 strain. The crude protein extract from E. coli BL21 carrying GST fusion protein or culture supernatant of B. subtilis carrying the chitinase gene exhibited enzyme activity by hydrolyzing chitin analogs, 4-methylumbelliferyl-$\beta$-D-N,N'-diacetylchitobioside and 4-methylumbelliferyl-$\beta$-D-N,N',N"-triacetylchitotrioside. These results indicated that even though the chitinase gene is not expressed in the N1 strain, the coding region is functional and encodes an active chitinase enzyme. Furthermore, B. subtilis 168 transformants expressing the chitinase gene exhibited antifungal activity against Fulvia fulva by suppressing spore germination. Our results suggest that the proper engineering of the expression of the indigenous chitinase gene, which will lead to its expression in the biocontrol strain B. licheniformis N1, may further enhance its biocontrol activity.

• Korean journal of applied entomology
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• v.53 no.3
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• pp.281-286
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• 2014

In grape vineyards, whitish spots in a cloud shape have been often observed on the fruit surface recently. However, the cause of the halo spot symptom was unknown, hindering countermeasures to be properly designed for the control. A small hole in the middle of the formless halo spot remained as a scar formed by oviposition of the thrips. It became later a suberized scab, which is separated from the epidermal cells on the surface either to be retained on or to be detached from it as time proceeds. Such a symptom is distinguished from the feeding damages caused by thrips or true bugs occurring on the grape fruits. With DNA extracted from the egg-shell found in the hole, molecular diagnosis by amplifying an ITS2 region with universal primers and subsequently digesting the PCR product by an restriction enzyme (RsaI) revealed that the egg was laid by Frankliniella occidentalis. In addition, a mitochondrial COI sequence confirmed that the halo spot symptom was formed by its oviposition. This study provides accurate information on the peculiar damage symptom caused by oviposition of F. occidentalis that could be useful in the control strategies for this pest in vineyards.

• Journal of Microbiology and Biotechnology
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• v.27 no.9
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• pp.1701-1710
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• 2017

Classical swine fever virus (CSFV) is the etiologic agent of classical swine fever, a highly contagious disease that causes significant economic losses to the swine industry. The lapinized C-strain, a widely used vaccine strain against CSFV, has low growth efficiency in cell culture, which limits the productivity in the vaccine industry. In this study, a recombinant virus derived from C-strain was constructed and subjected to continuous passaging in PK-15 cells with the goal of acquiring a high progeny virus yield. A cell-adapted virus variant, RecCpp80, had nearly 1,000-fold higher titer than its parent C-strain but lost the ability to induce fever in rabbits. Sequence analysis of cell-adapted RecC variants indicated that at least six nucleotide changes were fixed in RecCpp80. Further adaption of RecCpp80 variant in swine testicle cells led to a higher virus yield without additional mutations. Introduction of each of these residues into the wild-type RecC backbone showed that one mutation, M979R (T3310G), located in the C-terminal region of E2 might be closely related to the cell-adapted phenotype. Rabbit inoculation revealed that $RecCpp40_{+10}$ failed to induce fever in rabbits, whereas $RecCpp80_{+10}$ caused a fever response similar to the commercial C-strain vaccine. In conclusion, the C-strain can be adapted to cell culture by introducing specific mutations in its E2 protein. The mutations in RecCpp80 that led to the loss of fever response in rabbits require further investigation. Continuous passaging of the C-strain-based recombinant viruses in PK-15 cells could enhance its in vitro adaption. The non-synonymous mutations at 3310 and 3531 might play major roles in the enhanced capacity of general virus reproduction. Such findings may help design a modified C-strain for improved productivity of commercial vaccines at reduced production cost.

• Research in Plant Disease
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• v.18 no.2
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• pp.133-138
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• 2012

In November 2009, a powdery mildew on glossy abelia (Abelia ${\times}$ grandiflora) was found in Seogwipo, Jeju Island, Korea. Further survey in the southern part of Korea, e.g., Jeju, Busan, and Tongyeong confirmed occurrence of the disease. White colonies were present on leaves, young stems, and flowers, detracting from their beauty in landscape plantings. Severely infected lesions were discolored to red-purplish. Based on the morphological characteristics and analysis of rDNA, the fungus associated with the symptoms was identified as Erysiphe abeliicola U. Braun & S. Takam. This work provides the morphological feature of its anamorph for the first time, which is characterized by having multi-lobed hyphal appressoria and short foot-cells of conidiophores. Morphological characteristics of mature chasmothecia were consistent with the previous Japanese record of this species. The sequence of internal transcribed spacer region of ribosomal DNA obtained from a Korean sample showed that this species places in the section Microsphaera of the genus Erysiphe in phylogenetic position, corresponding with the classical taxonomy. This is the first report of E. abeliicola and its host plant in Korea. The host plant A. ${\times}$ grandiflora is newly listed in the host range of E. abeliicola.

• Korean Journal of Ichthyology
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• v.32 no.2
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• pp.39-48
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• 2020

The slender shinner (Pseudopungtungia tenuicorpa), a tiny freshwater fish of about 8 to 10 cm belonging to Cyprinidae, is an endangered species found only in the Han and Imjin Rivers on the Korean Peninsula. During the breeding season, this species spawns in nests of Coreoperca herzi, a predator of this species, or small crevices on rocks. This unique reproductive ecology can make this species more vulnerable to anthropogenic perturbance that can further limit the places to spawn. Here, mtDNA and microsatellite loci were analyzed to identify the genetic diversity and structure of slender shinners and further to provide the basic data necessary for the conservation planning of this species. A total of 28 polymorphic microsatellite markers were developed using Illumina paired-end sequencing, and 67 slender shinners collected from three localities in the Han River were genotyped using these loci. This species showed a remarkably high level of genetic diversity with mean expected heterozygosity of 0.914 and mean allele number per locus of 27.9, and no signature of drastic demographic decline was detected. As a result of our microsatellite analysis, the genetic structure between the two stems of the Han River, North Han and South Han, was prominent. Such a genetic structure was also evident in the sequence analysis of 14 haplotypes obtained from mtDNA control region. Although slender shinners are only found in very limited areas around the world, the genetic structure indicates that there is a block of gene flow among the populations, which should be reviewed in the future if management and restoration of this species is needed.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.5
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• pp.2073-2080
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• 2015

Background: The human papillomavirus (HPV) and its variants show wide geographical distribution and have been reported to cause cervical lesions. With cervical neoplasia as the leading cancer in Indian women, the aim of the present study was to evaluate the multiple infection HPV type distribution and variant genotypes in cervical samples from the coastal Karnataka region, India. Materials and Methods: A total of 212 samples were screened by nested polymerase chain reaction using PGMY9/11 and GP5+/6+ primers. HPV positive samples were sequenced to identify the types and a phylogenetic tree was constructed using the neighbor-joining method. Results: Sequence analysis identified a total of 14 HPV types distributed in 20%, 73.3% and 82.5% of non-malignant, pre-malignant [low grade squamous intraepithelial lesion (LSIL) and high grade squamous intraepithelial lesion (HSIL)] and cervical cancer samples. The distribution of high risk HPV in cancer samples was HPV 16, 76.4%, HPV18, 11.7%, HPV81, 2.9%, HPV31, 1.4%, HPV35, 1.4% and HPV 45, 1.4%. Multiple infections were observed in 11.8% of tumor samples with HPV 16 contributing to 62.5% of cases. In non-malignant samples, 20% of HPV positive samples were detected with HPV16, 82.3%, HPV33, 5.8% and HPV58, 5.8% and very low incidence of multiple infections. Comparative phylogenetic analysis of HPV variants identified 9 HPV sequences as new papillomavirus species, predominantly classified as European lineage type. Conclusions: The findings for HPV infections associated with progression of cervical cancer in coastal Karnataka region and HPV variant analysis provide baseline data for prevention and HPV vaccination programs.

• Journal of Microbiology and Biotechnology
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• v.12 no.1
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• pp.114-120
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• 2002

A 1,989-bp genomic region encoding nickel resistance genes was isolated from Legionella pneumophila, a pathogen for legionellosis. From a sequencing and computer analysis, the region was found to harbor two structural genes, a nreB-like protein gene (1,149 bp) and a nreA-like protein gene (270 bp), in a row. Both genes exhibited a significant degree of similarity to the corresponding genes from Synechocystis sp. PCC6803 ($54\%$ amino acid sequence identity) and Achromobacter xylosoxidans 31A ($76\%$). The gene was successfully expressed in E. coli MG1655 and conferred a nickel resistance of up to 5 mM in an LB medium and 3 mM in a TMS medium including gluconate as the sole carbon source. E. coli harboring the nickel resistance gene also exhibited a substantial resistance to cobalt, yet no resistance to cadmium or zinc. Since the extracellular concentration of nickel remained constant during the whole period of cultivation, it was confirmed that the nickel resistance was provided by an efflux system like the $Ni^2＋$permease (nrsD) of Synechocystis sp. strain PCC6803. Since polyphosphate (poly-P) is known as a global regulator for gene expression as well as a potential virulence factor in E. coli, the nickel resistance of a ppk mutant of E. coli MG 1655 harboring the nickel resistance gene from L. pneumophila was compared with that of its parental strain. The nickel resistance was significantly attenuated by ppk inactivation, which was more pronounced in an LB medium than in a TMS medium.

• Journal of Korea Multimedia Society
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• v.13 no.7
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• pp.1000-1015
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• 2010

In this paper, we propose a novel algorithm for hand gesture recognition. The hand detection method is based on human skin color, and we use the boundary energy information to locate the hand region accurately, then the moment method will be employed to locate the hand palm center. Hand gesture recognition can be separated into 2 step: firstly, the hand posture recognition: we employ the parallel NNs to deal with problem of hand posture recognition, pattern of a hand posture can be extracted by utilize the fitting ellipses method, which separates the detected hand region by 12 ellipses and calculates the white pixels rate in ellipse line. the pattern will be input to the NNs with 12 input nodes, the NNs contains 4 output nodes, each output node out a value within 0~1, the posture is then represented by composed of the 4 output codes. Secondly, the hand gesture tracking and recognition: we employed the Kalman filter to predict the position information of gesture to create the position sequence, distance relationship between positions will be used to confirm the gesture. The simulation have been performed on Windows XP to evaluate the efficiency of the algorithm, for recognizing the hand posture, we used 300 training images to train the recognizing machine and used 200 images to test the machine, the correct number is up to 194. And for testing the hand tracking recognition part, we make 1200 times gesture (each gesture 400 times), the total correct number is 1002 times. These results shows that the proposed gesture recognition algorithm can achieve an endurable job for detecting the hand and its' gesture.

• Korean Journal of Plant Taxonomy
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• v.37 no.1
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• pp.1-15
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• 2007

Phylogenetic studies were conducted for 42 populations of Korean viola based on matK gene and atpB-rbcL intergenic spacer region of chloroplast DNA. In the matK tree, section Chamaemelanium and Dischidium were formed as a distinct group. Five subsections of section Nomimium were paraphyletic. In atpB-rbcL intergenic spacer region analysis, two species of sect. Chamaemelanium were monophyletic, and section Dischidium was placed sister to subsection patellares clade except for V. keiskei. Five subsections of section Nomimium were also paraphyletic as matK tree. the separate data analyses were incongruent in the relationships among 42 populations, especially for the position of section Dischidium and V. keiskei. The combined analyses of two chloroplast regions showed three major clades; section Chamaemelanium and Dischidium (x=6) formed a sister to subsections Hypocarpae and Trigonocarpae (x=10) clade; subsections Bilobatae and vaginatae (x=10 or 12) formed a clade with V. keiskei; and 19 populations of subsection patellares (x=12) except for V. keiskei were recognized as an independent clade within section Nomimium. Although combined data suggest three major clades of Korean viola, the origins of each clade from outgroup were discordance with previous ITS and trnL-F data.

• Journal of Life Science
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• v.26 no.1
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• pp.1-7
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• 2016

Fatness is one of the most important economic traits in pigs. The leptin receptor (LEPR) gene may be a potential candidate for the fatness quantitative trait locus (QTL) on porcine chromosome 6, due to its position and physiological role. Thus, this study was carried out to evaluate the associations between structural variants in the LEPR gene and economic traits in pigs. We obtained an approximately 114-kb sequence containing the complete genomic DNA of the porcine LEPR gene, using shotgun sequencing of a bacterial artificial chromosome clone. We report the complete genomic structure of the porcine LEPR gene. Dozens of transcription factor-binding sites were found in the 1.2 kb upstream region from the transcription start point. An association study was performed with 550 Duroc pigs for 24 single-nucleotide polymorphisms (SNPs), including 6 SNPs within exons and 18 SNPs within the putative 5‘ regulatory region of the porcine LEPR gene. Among them, one SNP (−790C/G) was significantly associated with backfat thickness and lean meat percentage, whereas the others, including two SNPs with missense polymorphisms, had no effect on any phenotype. These results suggest that SNP −790C/G may be a useful marker for genetic improvements of fatness and leanness in Duroc pigs.