This study was carried out to investigate the expression patterns of foreign gene that controlled by tuber-specific patatin promoter in transgenic potatoes. Potato leaf disc cultured in vitro were transformed by the Agrobacterium strain LBA4404 containing pBl121 or pATGUS from potato cv. Desiree. In order to select the transgenic lines, gene-specific primers deduced from the NPTII were synthesized and used for polymerase chain reaction. The down part of the putative transgenic potatoes was transplanted weekly onto sucrose-enriched medium to accelerate the microtuber formation. RNA gel blot analysis was performed on the total RNAs obtained from tuber that had been harvested at a week interval. Also, histochemical assay was observed in the explants transformed with either pBI121 or pATGUS. Results showed that the transgenic plant containing pATGUS expressed GUS transcripts mainly at the tuber, not in stem, with the highest expression level in 5 weeks-grown microtubers. In contrast to pATGUS plants, the transformed plants with pBI121 showed an equal expression pattern throughout the whole developing stages. Consistent with RNA gel blot analysis, histochemical GUS staining and enzyme activity exhibited pATGUS transcripts were at the highest level in 5 weeks cultures. From these results, we suggest that the best stage to analyze the foreign gene introduced by patatin promoter into potato plants is at 5 weeks cultures after tuber formation.
This study analyzes the tide deformation of land boundary regions on the east (Region A) and west (Region B) sides of the Ross Ice Shelf in Antarctica using Double-Differential Interferometric Synthetic Aperture Radar (DDInSAR). A total of seven Sentinel-1A SAR images acquired in 2015-2016 were used to estimate the accuracy of tide prediction model and Young's modulus of ice shelf. First, we compared the Ross Sea Height-based Tidal Inverse (Ross_Inv) model, which is a representative tide prediction model for the Antarctic Ross Sea, with the tide deformation of the ice shelf extracted from the DDInSAR image. The accuracy was analyzed as 3.86 cm in the east region of Ross Ice Shelf and it was confirmed that the inverse barometric pressure effect must be corrected in the tide model. However, in the east, it is confirmed that the tide model may be inaccurate because a large error occurs even after correction of the atmospheric effect. In addition, the Young's modulus of the ice was calculated on the basis of the one-dimensional elastic beam model showing the correlation between the width of the hinge zone where the tide strain occurs and the ice thickness. For this purpose, the grounding line is defined as the line where the displacement caused by the tide appears in the DDInSAR image, and the hinge line is defined as the line to have the local maximum/minimum deformation, and the hinge zone as the area between the two lines. According to the one-dimensional elastic beam model assuming a semi-infinite plane, the width of the hinge region is directly proportional to the 0.75 power of the ice thickness. The width of the hinge zone was measured in the area where the ground line and the hinge line were close to the straight line shown in DDInSAR. The linear regression analysis with the 0.75 power of BEDMAP2 ice thickness estimated the Young's modulus of 1.77±0.73 GPa in the east and west of the Ross Ice Shelf. In this way, more accurate Young's modulus can be estimated by accumulating Sentinel-1 images in the future.
Yu, Yong-Man;Youn, Young-Nam;Hua, Quan Juan;Cha, Guang-Ho;Lee, Young-Ha
Journal of Food Hygiene and Safety
/
v.24
no.2
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pp.174-181
/
2009
The consumption of "ready-to-eat" agricultural products is recently increasing and the safety of these agricultural products is forefront of public concerns. The 120 samples of paprikas, strawberries and tomatoes, which are the representative exported agricultural products, were purchased at the department stores and discount stores in Daejeon. And we determined the microbiological and parasitological contamination level of these agricultural products using culture media, multiplex PCR, commercial bacterial detection kit and microscopy, and also evaluated the decontamination method. Mean counts of total aerobic bacteria from these agricultural products ranged from $1.3{\times}10^4$ CFU/g to $1.8{\times}10^5$ CFU/g, and mean counts of coliforms ranged from $1.4{\times}10^3$ CFU/g to $9.6{\times}10^3$ CFU/g. There was no significant difference in the level of bacterial contamination between the agricultural products from department stores and the ones from discount stores. Strawberry showed the highest contamination level for the bacteria and we also found the unidentified parasite eggs. Enterobacter cloacae was the most frequently isolated bacteria strain, but no food poisoning pathogenic bacteria except Staphylococcus aureus was isolated from the products by multiplex PCR. Compared to unwashed products, tab water-washed ones showed 80% decrease of the counts of total aerobic bacteria on the agricultural products, and the rates decreased more by incorporating detergent or ultrasonic wave treatment. We concluded that the biological contamination levels among paprikas, strawberries and tomatoes were the highest in strawberries, but there were not significant difference according to distribution systems.
Journal of Dental Rehabilitation and Applied Science
/
v.28
no.2
/
pp.147-161
/
2012
This study is to assess the effect of horizontal misfit of an implant supported 3-unit fixed prosthodontics on the stress development at the marginal cortical bone surrounding implant neck. Two finite element models consisting of a three unit fixed prosthodontics and an implant/bone complex were constructed on a three dimensional basis. The three unit fixed prosthodontics were designed either shorter (d=17.8mm model) or longer (d=18.0mm model) by 0.1mm than the span of two implants placed at the mandibular second premolar and second molar areas 17.9mm apart. Fitting of the fixed prosthodontics onto the implant abutments was simulated by a total of 6 steps, that is to say, 0.1mm displacement per each step, using DEFORM 3D (ver 6.1, SFTC, Columbus, OH, USA) program. Stresses in the fixed prosthodontics and implants were evaluated using von-Mises stress, maximum compressive stress, and radial stress as necessary. The d=17.8mm model assembled successfully on to the implant abutments while d=18.0mm model did not. Regardless if the fixed prosthodontics fitted onto the abutments or not, excessively higher stresses developed during the course of assembly trial and thereafter. On the marginal cortical bone around implants during the assembly, the peak tensile and compressive stresses were as high as 186.9MPa and 114.1MPa, respectively, even after the final sitting of the fixed prosthodontics (for d=17.8mm model). For this case, the area of marginal bone subject to compressive stresses above 55MPa, equivalent of the $4,000{\mu}{\varepsilon}$, i.e. the reported threshold strain to inhibit physiological remodeling of human cortical bone, extended up to 2mm away from implant during the assembly. Horizontal misfit of 0.1mm can produce excessively high stresses on the marginal cortical bone not only during the fixed prosthodontics assembly but also thereafter.
Pseudomonas aeruginosa is a Gram (-) opportunistic human pathogen causing a wide variety of infections on lung, urinary tract, eyes, and burn wound sites and quorum sensing (QS), a cell density-sensing mechanism plays an essential role in Pseudomonas pathogenesis. In order to investigate the importance of QS in the Pseudomonas infections of Korean patients, we isolated 189 clinical strains of P. aeruginosa from the patients in Pusan Paik Hospital, Busan, South Korea. The QS signal production of these clinical isolates was measured by signal diffusion assay on solid media using reporter strains. While most clinical strains (79.4%) produced the QS signals as similar level as a wild type strain, PAO1 did, where LasR, the initial QS signal sensor-regulator was fully activated, a minority of them (4.2%) produced much less QS signals at the level to which LasR failed to respond. Similarly, while 72.5% of the clinical isolates produced QS signals enough to activate QscR, an another QS signal sensor-regulator, some few of them (9%) produced the QS signals at much lower level where QscR was not activated. For further analysis, we selected 74 clinical strains that were obtained from the patients under suspicion of Pseudomonas infection and investigated the total protease activity that is considered important for virulence. Interestingly, significant portion of them showed very low protease activity (44.6%) or no detectable protease activity (12.2%). When the biofilm-forming ability that is considered very important in chronic infection was examined, most isolates showed lower biofilm-forming activity than PAO1. Similarly, significant portion of clinical isolates showed reduced motility (reduced swarming activity in 51.4% and reduced twitching activity in 41.9%), or non-detectable motility (swarming-negative in 28.4% and twitching-negative in 28.4%). Our result showed that the clinical isolates that produced QS signals at the similar level to wild type could have significantly reduced activities in the protease production, biofilm formation, and motility, and some clinical isolates had unique patterns of motility, biofilm formation, and protease production that are not correlated to their QS activity.
Rice flour is used in many food products. However, dough made from rice lacks extensibility and elasticity, making it less suitable than wheat for many food products such as bread and noodles. The high-molecular weight glutenin subunits (HMW-GS) of wheat play a crucial role in determining the processing properties of the wheat grain. This paper describes the development of marker-free transgenic rice plants expressing a wheat Glu-Dy10 gene encoding the HMG-GS from the Korean wheat cultivar 'Jokyeong' using Agrobacterium-mediated co-transformation. Two expression cassettes, consisting of separate DNA fragments containing Glu-1Dy10 and hygromycin phosphotransferase II (HPTII) resistance genes, were introduced separately into Agrobacterium tumefaciens EHA105 for co-infection. Each EHA105 strain harboring Glu-1Dy10 or HPTII was infected into rice calli at a 3: 1 ratio of Glu-1Bx7 and HPTII. Among 290 hygromycin-resistant $T_0$ plants, we obtained 29 transgenic lines with both the Glu-1Dy10 and HPTII genes inserted into the rice genome. We reconfirmed the integration of the Glu-1Dy10 gene into the rice genome by Southern blot analysis. Transcripts and proteins of the Glu-1Dy10 in transgenic rice seeds were examined by semi-quantitative RT-PCR and Western blot analysis. The marker-free plants containing only the Glu-1Dy10 gene were successfully screened in the $T_1$ generation.
Sam Woong Kim;Young Jin Kim;Tae Wook Lee;Won-Jae Chi;Woo Young Bang;Tae Wan Kim;Kyu Ho Bang;Sang Wan Gal
Journal of Life Science
/
v.33
no.11
/
pp.897-904
/
2023
This study was done to develope genetic markers with the unique characteristics of genes according to the genomic information of Bacillus velezensis K10. B. velezensis K10 maintained a total of 4,159,835 bps, which was found to encode 5,136 open reading frames (orfs). B. velezensis K10 was found to have much more gene migration due to external factors overall compared to standard strain B. velezensis JS25R. In order to discover genetic selection markers, orfs on the genome to be easily induced to gene mutation were surveyed such as recombinase, integrase, transposase, and phage-related genes. As a result of the investigation, 9 candidate markers were isolated with high possibility as genetic selection markers. Although a part in the various origin's areas showed specificities in comparison with homology, the selected markers were all existed in phage-related areas because they were relatively lower homologies in phage-related genes. PCR analysis was done on B. licheniformis K12, B. velezensis K10, B. subtilis, and B. cereus to establish them as inter-species candidate selection markers. As a result, it was confirmed that B. velezensis K10-specific PCR products were formed in a total of 6 primer sets such as BV3 and BV5 to 9. On the other hand, analysis at the subspecies level observed the formation of B. velezensis K10-specific PCR products in 4 primer sets such as BV3, 5, 8, and 9. Among them, since BV5 and BV8 were detected by very specific results, we suggest that BV5 and 8 can be used as B. velezensis K10 gene selection markers at the species and sub-species level.
Sam Woong Kim;Young Jin Kim;Hyo In Choi;Sang Won Lee;Won-Jae Chi;Woo Young Bang;Tae Wan Kim;Kyu Ho Bang;Sang Wan Gal
Journal of Life Science
/
v.34
no.7
/
pp.465-475
/
2024
Lactiplantibacillus plantarum K9 is a probiotic strain that can be utilized from various bioactive substances isolated from Protaetia brevitarsis seulensis larvae. In this study, a genetic analysis of L. plantarum K9 revealed the existence of a bacterial chromosome and three plasmids. The glycolysis pathway and pentose phosphate pathway were examined for their normal functioning via an analysis of the core metabolic pathways of L. plantarum K9. Since the key enzymes, fluctose-1,6-bisphospatase (EC: 3.1.3.11) and 6-phosphogluconate dehydratase (EC: 4.2.1.12)/2-keto-deoxy-6-phosphogluconate (KDPG) aldolase (EC: 4.2.1.55), of gluconeogenesis and the ED pathway were not identified from the L. plantarum K9 genome, we suggest that gluconeogenesis and the ED pathway are not performed in L. plantarum K9. Additionally, while some enzymes, related to fumarate and malate biosyntheses, involved in the TCA cycle were identified from L. plantarum K9, the enzymes associated with the remaining TCA cycle were absent, indicating that the TCA cycle cannot proceed. Meanwhile, based on our findings, we propose that the oxidative electron transport system performs class IIB-type (bd-type) electron transfer. In summary, we assert that L. plantarum K9 performs homolactic fermentation, executes gluconeogenesis and the pentose phosphate pathway, and carries out energy metabolism through the class IIB-type oxidative electron transport system. Therefore, we suggest that L. plantarum K9 has relatively high lactic acid production, and that it has excellent antibacterial activity, as a result, compared to other lactic acid bacterial strains. Moreover, we speculate that L. plantarum K9 has an oxidative electron transport capability, indicating that it is highly resistant to oxygen and suggesting that it has fine cultivation characteristics, which collectively make it highly suitable for use as a probiotic.
A fungal strain of Trichoderma having strong chitinolytic activity was isolated from field soil enriched with crabshell for several years. Based on 5.8S rRNA, partial 18S, 28S rRNA genes, ITS1, ITS2 sequence analysis and morphological characteristics, the fungus was identified as Trichoderma asperellum and named as Trichoderma asperellum T-5 (TaT-5). The fungus released lytic enzymes such as chitinase and ${\beta}$-1, 3-glucanse, and produced six antifungal substances in chitin broth medium. To demonstrate the protective effect of TaT-5 against damping-off in cucumber plant caused by Rhizoctonia solani, TaT-5 culture broth (TA), chitin medium (CM) and distilled water (DW) were applied to each pot at 10 days after sowing, respectively. Then, the homogenized hyphae of R. solani were infected to each pot at 1 week after TaT-5 inoculation. During experimental period, fresh weight of shoot and root in cucumber plant more increased at TA treatment compared to other treatments. PR-proteins (${\beta}$-1, 3-glucanase and chitinase) activities in cucumber leaves markedly increased at CM and DW treatments, but the activity slightly increased and then decreased at TA treatment at 3 days after infection of R. solani. The activity of PR-proteins activities in cucumber roots at all treatments decreased with time where the degree of decrement was more alleviated at TA treatment than CM and DW. These results suggest that the lytic enzymes (chitinase and ${\beta}$-1, 3-glucanse) and antifungal substances produced by TaT-5 can reduce the pathogenic attack by R. solani in cucumber plants.
Journal of the Microelectronics and Packaging Society
/
v.25
no.1
/
pp.1-10
/
2018
Recently, there has been rapid development in the field of flexible electronic devices, such as organic light emitting diodes (OLEDs), organic solar cells and flexible sensors. Encapsulation process is added to protect the flexible electronic devices from exposure to oxygen and moisture in the air. Using numerical simulation, we investigated the effects of the encapsulation layer on mechanical stability of the silicon chip, especially the fracture performance of center crack in multi-layer package for various loading condition. The multi-layer package is categorized in two type - a wide chip model in which the chip has a large width and encapsulation layer covers only the chip, and a narrow chip model in which the chip covers both the substrate and the chip with smaller width than the substrate. In the wide chip model where the external load acts directly on the chip, the encapsulation layer with high stiffness enhanced the crack resistance of the film chip as the thickness of the encapsulation layer increased regardless of loading conditions. In contrast, the encapsulation layer with high stiffness reduced the crack resistance of the film chip in the narrow chip model for the case of external tensile strain loading. This is because the external load is transferred to the chip through the encapsulation layer and the small load acts on the chip for the weak encapsulation layer in the narrow chip model. When the bending moment acts on the narrow model, thin encapsulation layer and thick encapsulation layer show the opposite results since the neutral axis is moving toward the chip with a crack and load acting on chip decreases consequently as the thickness of encapsulation layer increases. The present study is expected to provide practical design guidance to enhance the durability and fracture performance of the silicon chip in the multilayer package with encapsulation layer.
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