• Title/Summary/Keyword: replication control

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Construction of a Novel Shuttle Vector for Tetragenococcus species based on a Cryptic Plasmid from Tetragenococcus halophilus

  • Min Jae Kim;Tae Jin Kim;Yun Ji Kang;Ji Yeon Yoo;Jeong Hwan Kim
    • Journal of Microbiology and Biotechnology
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    • v.33 no.2
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    • pp.211-218
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    • 2023
  • A cryptic plasmid (pTH32) was characterized from Tetragenococcus halophilus 32, an isolate from jeotgal, Korean traditional fermented seafood. pTH32 is 3,198 bp in size with G+C content of 35.84%, and contains 4 open reading frames (ORFs). orf1 and orf2 are 456 bp and 273 bp in size, respectively, and their translation products showed 65.16% and 69.35% similarities with RepB family plasmid replication initiators, respectively, suggesting the rolling-circle replication (RCR) mode of pTH32. orf3 and orf4 encodes putative hypothetical protein of 186 and 76 amino acids, respectively. A novel Tetragenococcus-Escherichia coli shuttle vector, pMJ32E (7.3 kb, Emr), was constructed by ligation of pTH32 with pBluescript II KS(+) and an erythromycin resistance gene (ErmC). pMJ32E successfully replicated in Enterococcus faecalis 29212 and T. halophilus 31 but not in other LAB species. A pepA gene, encoding aminopeptidase A (PepA) from T. halophilus CY54, was successfully expressed in T. halophilus 31 using pMJ32E. The transformant (TF) showed higher PepA activity (49.8 U/mg protein) than T. halophilus 31 cell (control). When T. halophilus 31 TF was subculturd in MRS broth without antibiotic at 48 h intervals, 53.8% of cells retained pMJ32E after 96 h, and only 2.4% of cells retained pMJ32E after 14 days, supporting the RCR mode of pTH32. pMJ32E could be useful for the genetic engineering of Tetragenococcus and Enterococcus species.

Multiple Alternating Immunizations with DNA Vaccine and Replication-incompetent Adenovirus Expressing gB of Pseudorabies Virus Protect Animals Against Lethal Virus Challenge

  • Kim, Seon-Ju;Kim, Hye-Kyung;Han, Young-Woo;Aleyas, Abi G.;George, Junu A.;Yoon, Hyun-A;Yoo, Dong-Jin;Kim, Koan-Hoi;Eo, Seong-Kug
    • Journal of Microbiology and Biotechnology
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    • v.18 no.7
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    • pp.1326-1334
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    • 2008
  • The prime-boost vaccination with DNA vaccine and recombinant viral vector has emerged as an effective prophylactic strategy to control infectious diseases. Here, we compared the protective immunities induced by multiple alternating immunizations with DNA vaccine (pCIgB) and replication-incompetent adenovirus (Ad-gB) expressing glycoprotein gB of pseudorabies virus (PrV). The platform of pCIgB-prime and Ad-gB-boost induced the most effective immune responses and provided protection against virulent PrV infection. However, priming with pCIgB prior to vaccinating animals by the DNA vaccine-prime and Ad-boost protocol provided neither effective immune responses nor protection against PrV. Similarly, boosting with Ad-gB following immunization with DNA vaccine-prime and Ad-boost showed no significant responses. Moreover, whereas the administration of Ad-gB for primary immunization induced Th2-type-biased immunity, priming with pCIgB induced Th1-type-biased immunity, as judged by the production of PrV-specific IgG isotypes and cytokine IFN-$\gamma$. These results indicate that the order and injection frequency of vaccine vehicles used for heterologous prime-boost vaccination affect the magnitude and nature of the immunity. Therefore, our demonstration implies that the prime-boost protocol should be carefully considered and selected to induce the desired immune responses.

Assessment of the resistance of bacterial soft rot caused by Pectobacterium carotovorum subsp. carotovorum KACC 21701 in Kimchi cabbage genetic resources

  • Parthiban Subramanian;Ho Chul Ko;Seong-Hoon Kim;Jae Eun Lee;Aejin Hwang;Bichsaem Kim;Yoon-Jung Lee;Awraris Derbie Assefa;Onsook Hur;Nayoung Ro;Jung Sook Sung;Ju Hee Rhee;Ho-Sun Lee;Bum-Soo Hahn
    • Korean Journal of Environmental Biology
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    • v.40 no.4
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    • pp.433-441
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    • 2022
  • Bacterial phytopathogen Pectobacterium causes soft rot disease in several vegetable crops globally, resulting in heavy agricultural losses at both the pre and postharvest stages. The present work was carried out to screen Kimchi cabbage genetic resources conserved at the National Agrobiodiversity Center, Rural Development Administration, Korea, for resistance against the soft rot pathogen Pectobacterium carotovorum subsp. carotovorum KACC 21701 over a period of three years (from 2020 to 2022). Infection of the phytopathogen was carried out at four-leaf stage and for each accession, twenty-five plants per germplasm were infected with KACC 21701. Kimchi cabbage cultivars Wangmatbaechu, Seoulbaechu, and CR Kiyoshi were used as control. Seven-days post-infection, the Disease Index (DI) values were manually recorded from zero to four, zero matched perfectly heathy plants and four completely dead plants. The 682 accessions of Kimchi cabbage exhibited varying degrees of disease resistance to KACC 21701 and thirty accessions, exhibiting a DI≤2, were considered for replication studies. During the replication studies, four landrace germplasms (IT102883, IT120036, IT120044, and IT120048) and one cultivar(IT187919) were confirmed to be moderately susceptible to KACC 21701. Results of the preliminary screening as well as replication studies were documented for the all the 682 germplasms. Addition of such information to the passport data of stored germplasms might serve as potential bio-resource for future breeders and researchers to develop resistant varieties or study the mechanisms involved in resistance of plants to such phytopathogen.

A Distributed Real-Time Concurrency Control Scheme using Transaction the Rise of Priority (트랜잭션 우선 순위 상승을 이용한 분산 실시간 병행수행제어 기법)

  • Lee, Jong-Sul;Shin, Jae-Ryong;Cho, Ki-Hyung;Yoo, Jae-Soo
    • Journal of KIISE:Databases
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    • v.28 no.3
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    • pp.484-493
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    • 2001
  • As real-time database systems are extended to the distributed computing environment, the need to apply the existing real-time concurrency control schemes to the distributed computing environment has been made. In this paper we propose an efficient concurrency control scheme for distributed real-time database system. Our proposed scheme guarantees a transaction to commit at its maximum, reduces the restart of a transaction that is on the prepared commit phase, and minimizes the time of the lock holding. This is because it raises the priority of the transaction that is on the prepared commit phase in the distributed real-time computing environment. In addition, it reduces the waiting time of a transaction that owns borrowed data and improves the performance of the system, as a result of lending the data that the transaction with the raised priority holds. We compare the proposed scheme with DO2PL_PA(Distributed Optimistic Two-Phase Locking) and MIRROR(Managing Isolation in Replicated Real-time Object Repositories) protocol in terms of the arrival rate of transactions, the size of transactions, the write probability of transactions, and the replication degree of data in a firm-deadline real-time database system based on two-phase commit protocol. It is shown through the performance evaluation that our scheme outperforms the existing schemes.

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Effects of Nitrogen Addition on Soil Respiration (상수리나무림 임상에 공급한 무기질소가 토양호흡에 미치는 영향)

  • 최주섭;문형태
    • The Korean Journal of Ecology
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    • v.27 no.3
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    • pp.155-159
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    • 2004
  • In order to gain a better understanding of how forests participate in the cycling of carbon, effects of nitrogen addition on soil respiration were investigated on the oak forest in Kongju, Korea. Study site was divided into control, treatment f and treatment 2 plots, with 5 replication in each plot. In each replicate of treatment 1 and treatment 2 were fertilized with ammonium nitrate (NH$_4$NO$_3$), 30 g/$m^2$ and 60 g/$m^2$, respectively. Soil respiration, soil temperature, ammonium-N and nitrate-N were measured during the experimental period. Ammonium-N and nitrate-N in Ta were higher than those in control and T$_1$. Ammonium-N and nitrate-N in top-soil and sub-soil decreased sharply in August after bi9 rainfall in July in T$_1$ and T$_2$, however, those in control plot increased. Soil respiration in T$_2$ Plot showed consistently higher than those in control and T$_1$ until the end of July. However, soil respiration was similar among the control, T$_1$ and T$_2$ in mid-August and September The amount of Co$_2$ released from soil respiration in control, T$_1$ and T$_2$ in mid-August was 8.0$\pm$0.4, 9.3$\pm$0.6 and 10.2$\pm$0.5 $\mu$mol$^{-1}$ ㆍm$^{-2}$ ㆍs$^{-1}$ , respectively. However, those in control, T$_1$ and T$_2$in mid-August was 13.0$\pm$0.4, 13.5$\pm$0.5, 13.3$\pm$0.6 $\mu$mol$^{-1}$ ㆍm$^{-2}$$^{-1}$ , respectively. The results suggest that nitrogen addition in this oak forest has a positive effect on soil respiration.

Association of an Anti-inflammatory Cytokine Gene IL4 Polymorphism with the Risk of Type 2 Diabetes Mellitus in Korean Populations

  • Go, Min-Jin;Min, Hae-Sook;Lee, Jong-Young;Kim, Sung-Soo;Kim, Yeon-Jung
    • Genomics & Informatics
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    • v.9 no.3
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    • pp.114-120
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    • 2011
  • Chronic inflammation has been implicated as one of the important etiological factors in insulin resistance and type 2 diabetes mellitus (T2DM). To investigate the role of anti-inflammatory cytokines in the development of T2DM, we conducted a case-control study to assess the association between IL4/IL4R polymorphisms and disease risk. We firstly identified single nucleotide poly-morphisms (SNP) at IL4 and IL4RA loci by sequencing the loci in Korean participants. Case-control studies were conducted by genotyping the SNPs in 474 T2DM cases and 470 non-diabetic controls recruited from community-based cohorts. Replication of the associated signals was performed in 1,216 cases and 1,352 controls. We assessed effect of IL4 -IL4RA interaction on T2DM using logistic regression method. The functional relevance of the SNP associated with disease risk was determined using a reporter expression assay. We identified a strong association between the IL4 promoter variant rs2243250 and T2DM risk (OR=0.77; 95% CI, 0.67~0.88; p=$1.65{\times}10^{-4}$ in the meta-analysis). The reporter gene expression assay demonstrated that the presence of rs2243250 might affect the gene expression level with ~1.5-fold allele difference. Our findings contribute to the identification of IL4 as a T2D susceptibility locus, further supporting the role of anti-inflammatory cytokines in T2DM disease development.

Effects of Feeding Dried Leftover Food on Growth and Body Composition of Broiler Chicks

  • Cho, Y.M.;Lee, G.W.;Jang, J.S.;Shin, I.S.;Myung, K.H.;Choi, K.S.;Bae, I.H.;Yang, C.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.3
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    • pp.386-393
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    • 2004
  • This study was conducted to investigate the effects of feeding dried leftover food (DLF) on growth, body composition and feed conversion of broiler chicks. One hundred ninety-six of one-day old Ross broiler chicks were assigned to 7 treatments in a completely randomized design. Each treatment had four replications with seven chicks per replication. The treatments groups included control without DLF, dietary 10% level of DLF, dietary 20% level of DLF and dietary 30% level of DLF, 5% higher protein level of diet containing 10% DLF, 10% higher protein level of diet containing 20% DLF and 15% higher protein level of diet containing 30% DLF than control diet. Body weight gain was slightly higher in control group than that of DLF-fed groups. However, there were no significant differences in body weight gain among those groups fed diets containing different levels of DLF. In general, increasing dietary level of DLF resulted in decreasing feed conversion. Content of crude protein in whole broiler body was slightly higher in control group although any significant difference was not found among treatments (p>0.05). Content of crude fat in whole broiler body was lowest in groups fed diets containing 30% DLF with 15% higher protein level than control diet, showing significant difference from groups fed diets containing 20% DLF (p<0.05). Contents of total cholesterol, free cholesterol, cholesterol ester and LDL- cholesterol in blood of broilers fed DLF-containing diets generally appeared to be higher compared with control group without significant difference (p>0.05). Fatty acid contents in broiler meat were higher in the order of oleic acid, palmitic acid and linoleic acid without significant differences among treatments. Content of DHA in broiler meat was higher in groups fed diets containing DLF than that of control group although there were no significant differences among treatments (p>0.05).

Effects of Feeding Dried Leftover Food on Productivity of Laying Hens

  • Cho, Y.M.;Shin, I.S.;Yang, C.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.4
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    • pp.518-522
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    • 2004
  • This experiment was conducted to investigate the effects of different levels of dried leftover food (DLF) in the diet on feed utilization and egg-laying performance of hens. One hundred sixty-eight, 18 week old Tetra brown commercial layers, were assigned to 7 treatments in a completely randomized design. Each treatment has four replications per treatment with six animals per replication. All the experimental animals were fed diets for 7 weeks. The treatments included 1) control group without DLF, 2) diet with 10% DLF, 3) diet with 20% DLF, 4) diet with 30% DLF, 5) 10% higher protein level of diet with 10% DLF, 6) 20% higher protein level of diet with 20% DLF and 7) 30% higher protein level of diet with 30% DLF. Average daily feed intake (ADFI) tended to be improved with DLF feeding. ADFI of group fed diets with 20% was significantly higher than that of control (p<0.05). Feed conversions of DLFfed groups were higher than that of control. Egg production tended to be higher in groups fed diets with 10% DLF than control diet without significant differences (p>0.05). However, those of groups fed diets containing 20 and 30% DLF were lower than that of control. Supplementing protein source to DLF-containing diets improved egg production (p<0.05). Increasing level of DLF in the diet for layer decreased egg weight and egg mass compared to control without significant differences (p>0.05). Protein supplementation to DLF-containing diets increased egg mass without significant difference (p>0.05). The range of egg cholesterol concentration of DLF-fed groups was 11.94-14.10 mg/g while that of control group was 12.31 mg/g although there was no significant difference among treatments (p>0.05).

Effects of Herbicide Application Method on Abutilon avicennar Control and Corn Yield (제초제 살포방법이 어저귀 방제 및 옥수수의 생산성에 미치는 영향)

  • Kang, W.S.;Kim, J.G.;Chung, E.S.;Seo, S.;Yang, J.S.
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.18 no.2
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    • pp.107-112
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    • 1998
  • This experiment was carried out to investigate effects of herbicide application method on Abutilon avicennae control and corn yield at the forage experimental field, grassland and forage crop division, National Livestock Research Institute, RDA, Suwon from 1996 to 1997. The experiment was arranged in a randomized block design with three replication. The herbicide application consisted of control, 2-3 leaf stage(Dicamba), 5-6 leaf stage (Dicamba) and soil treatment(Pendimetha1in). The hybrid of corn used in this experiment was P 3352. The results obtained are summarized as follow; 1. The plant and ear height was the lowest at control. Tasseling and silking date were delayed 3-4 day at control. The length and weight of ear were the highest at Dicamba treatment in 2-3 leaf stage. 2. The herbicide injury of corn was detected slightly after 10-20 day but the injury was recovered soon. The weed control efficiency was 96.7 and 81.8% at Dicamba treatment in 2-3 and 5-6 leaf stage, respectively. 3. The fresh and dry matter yield of Dicamba treatment in 2-3 leaf stage was higher than that of control by 36%, but no significant difference was found among herbicide application method. 4. Herbicide application method had little effect on the ADF, NDF and CP contents, but the CP and IVDMD of ear were higher than that of stover. The results of this experiment indicate that the application of Dicarnba in 2-6 leaf stage of corn will remove almost all of A butilon avicennae and increase DM yield of corn.

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Alternation to the Randomized Block Design for Agricultural Experiments in Korea (농업실험에서 임의화블록설계에 대한 대안 - 농촌진흥청 사례들을 중심으로 -)

  • 허명회;한원식;신한풍
    • The Korean Journal of Applied Statistics
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    • v.10 no.1
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    • pp.15-27
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    • 1997
  • Randomized block design (RBD) with three replication is very frequently adopted in agricultural experiments of the Rural Development Administration of Korea. Even though it works well in field trials of traditional crops, it may not accomodate trial site conditions and/or experimental environment. In this research report, we deal with two such cases. The first case is for a crop experiment in green houses. In house conditions, RBD may not be appropriate since it cannot reflect two directions of the yield gradient. So, a Latin square design is suggested as an alternative. The second case is for local field experiments of the newly-inbred rice. RBD with three replications is used without doubt for decades, even though the site layout is not appropriately shaped for the design. In this case, we suggest the RBD in two blocks with multiple replicates for control varieties as an alternative. To improve the quality of statistical experimental designs in over one-thousand agricultural trials performed annually in the Rural Development Administration, we need to re-train agricultural researchers on the design and analysis of experiments and call for concerns of Korean statisticians.

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