• Journal of Microbiology and Biotechnology
• /
• v.21 no.4
• /
• pp.374-378
• /
• 2011

To find alternative genetic resources for D-serine dehydratase (E.C. 4.3.1.18, dsdA) mediating the deamination of D-serine into pyruvate, metagenomic libraries were screened. The chromosomal dsdA gene of a wild-type Escherichia coli W3110 strain was disrupted by inserting the tetracycline resistance gene (tet), using double-crossover, for use as a screening host. The W3110 dsdA::tet strain was not able to grow in a medium containing D-serine as a sole carbon source, whereas wild-type W3110 and the complement W3110 dsdA::tet strain containing a dsdA-expression plasmid were able to grow. After introducing metagenome libraries into the screening host, a strain containing a 40-kb DNA fragment obtained from the metagenomic souce derived from a compost was selected based on its capability to grow on the agar plate containing D-serine as a sole carbon source. For identification of the genetic resource responsible for the D-serine degrading capability, transposon-${\mu}$ was randomly inserted into the 40-kb metagenome. Two strains that had lost their D-serine degrading ability were negatively selected, and the two 6-kb contigs responsible for the D-serine degrading capability were sequenced and deposited (GenBank code: HQ829474.1 and HQ829475.1). Therefore, new alternative genetic resources for D-serine dehydratase was found from the metagenomic resource, and the corresponding ORFs are discussed.

• The Journal of the Korea Contents Association
• /
• v.3 no.2
• /
• pp.65-72
• /
• 2003

In this paper, the new scheme using a Combined Cache(CC) that combing the Paging and Routing Cache(PRC) and an alternative handoff method according to the type of data transmission for achieving the efficient realtime communication. The PRC and quasi-soft handoff method reduce the path duplication. But they increase the network traffic load because of the handoff state packet of Mobile Host(MH). Moreover the use the same handoff method without differentiating the type of transmission data. Those problems are solved by operating U with a semi-soft handoff method for realtime data transmission and with a hard handoff method for non-realtime data transmission. As a result or simulation a better performance is obtained because of the reduction of the number of control packet in case that the number of cells are below 20. And the packet arrival time and loss of packet decreased significantly for realtime data transmission.

• Journal of Microbiology and Biotechnology
• /
• v.33 no.3
• /
• pp.329-338
• /
• 2023

Enterohemorrhagic Escherichia coli (EHEC) is a foodborne pathogen that produces attaching and effacing lesions on the large intestine and causes hemorrhagic colitis. It is primarily transmitted through the consumption of contaminated meat or fresh produce. Similar to other bacterial pathogens, antibiotic resistance is of concern for EHEC. Furthermore, since the production of Shiga toxin by this pathogen is enhanced after antibiotic treatment, alternative agents that control EHEC are necessary. This study aimed to discover alternative treatments that target virulence factors and reduce EHEC toxicity. The locus of enterocyte effacement (LEE) is essential for EHEC attachment to host cells and virulence, and most of the LEE genes are positively regulated by the transcriptional regulator, Ler. GrlA protein, a transcriptional activator of ler, is thus a potential target for virulence inhibitors of EHEC. To identify the GrlA inhibitors, an in vivo high-throughput screening (HTS) system consisting of a GrlA-expressing plasmid and a reporter plasmid was constructed. Since the reporter luminescence gene was fused to the ler promoter, the bioluminescence would decrease if inhibitors affected the GrlA. By screening 8,201 compounds from the Korea Chemical Bank, we identified a novel GrlA inhibitor named Grlactin [3-[(2,4-dichlorophenoxy)methyl]-4-(3-methylbut-2-en-1-yl)-4,5-dihydro-1,2,4-oxadiazol-5-one], which suppresses the expression of LEE genes. Grlactin significantly diminished the adhesion of EHEC strain EDL933 to human epithelial cells without inhibiting bacterial growth. These findings suggest that the developed screening system was effective at identifying GrlA inhibitors, and Grlactin has potential for use as a novel anti-adhesion agent for EHEC while reducing the incidence of resistance.

• Journal of Life Science
• /
• v.11 no.4
• /
• pp.385-391
• /
• 2001

Filamentous hemagglutinin (FHA) is considered as an essential immunogenic component for incorporation into acellular vaccines against Bordetella pertussis, the causative agent of whooping cough. Classically, antipertussis vaccination has employed an intramuscular route. An alternative approach to stimulate mucosal and systemic immune responses is oral immunization with recombinant live vaccine carrier strains of Salmonella typhimurium. An attenuated live Salmonella vaccine sgrain($\Delta$cya $\Delta$crp) expressing recombinant FHA(rFHA) was developed. Stable expressionof rFHA was achieved by the use of balanced-lethal vector-host system. which employs an asd deletion in the host chromosome to impose in obligate requirement for diaminopimelic acid. The chromosomal $\Delta$asd mutation was complemented by a plasmid vector possessing the asd$^{+}$ gene. A 3 kb DNA fragment encoding immuno dominant regionof FHA was subcloned in-frame downstream to the ATG translation initiation codon in the multicopy Asd$^{+}$ pYA3341 vector to create pYA3457. Salmonella vaccine harboring pYA3457 expressed approximately 105kDa rFHA protein. The 100% maintenance of [YA3457 in vaccine strain was confirmed by stability examinations. Additionally, a recombinant plasmid pYA3458 was constructed to overpress His(8X)-tagged rFHA in Essherichia coli. His-tagged rFHA was purified from the E. coli strain harboring pYA3458 using Ni$^{2+}$-NTA affinity purification system.>$^{2+}$-NTA affinity purification system.

• Molecules and Cells
• /
• v.25 no.3
• /
• pp.446-451
• /
• 2008

We assessed heterologous protein expression in 64 strains obtained from the Escherichia coli Reference (ECOR) collection, a collection representing diverse natural E. coli populations. A plasmid generating a glutathione S-transferase and plant carbonic anhydrase fusion protein (GST-CA) under the control of the tac promoter was introduced into the ECOR strains, and the quantity of the fusion protein was determined by SDS-PAGE. The foreign protein was generated at various levels, from very high (40 strains, high producers) to very low (six strains, low producers). Immunoblotting showed that the high producers expressed approximately 250-500 times more GST-CA protein than the low producers. The results of semi-quantitative RT-PCR showed that the low producers generated mRNA levels comparable to those of the high producers, thereby suggesting that, at least in this case, inefficient translation is a major cause of the low production. We introduced a different plasmid, which expressed a maltose binding protein and plant guanylate kinase fusion protein (MBP-GK) into the six low producers. Interestingly, five of these expressed MBP-GK at very high levels. Thus, we conclude that the production of a particular protein from an expression vector can vary considerably, depending on the host strain. Strains in the ECOR collection could function as useful alternative hosts when a desired level of protein expression is not obtained from commonly used strains, such as E. coli K12 or B derivatives.

• The Journal of the Korea institute of electronic communication sciences
• /
• v.17 no.5
• /
• pp.783-790
• /
• 2022

The development of SNS and video platforms provided an opportunity to explode the activation of content production and consumption. However, in the legacy system, due to the host-based location-oriented data transmission, there are inherent limitations in efficient operation and management. As an alternative to this, a Contents Centric Network (CCN) was studied. In this paper, when intermediate nodes located between the information provider and the information requester between the real-time streaming services in the CCN environment move or restrict their use, failure through monitoring of wireless reception strength to solve problems like disconnection of transmission quality at the information consumer. We propose a stable intermediate node management mechanism through active response before occurrence.

• Korean journal of applied entomology
• /
• v.55 no.4
• /
• pp.389-403
• /
• 2016

Sugar beet nematode (Heterodera schachtii Schmidt) was first detected in 2011, in Chinese cabbage grown in the highland areas of Korea. Chemical control of the nematode by nematicides is not feasible due to its cyst-forming characteristics; therefore, the cultivation of non-host crops is a preferable alternative to utilize nematode-infected fields. In this study, a total of 276 plant cultivars belonging to 18 different families were screened to evaluate their resistance to the nematode. Based on the number of cysts formed following nematode inoculation, the tested crops were classified into 3 levels: susceptible, moderately susceptible, and resistant/immune. Among the 276 cultivars tested, 106 cultivars were susceptible, 40 cultivars were moderately susceptible, and 130 cultivars were resistant/immune. Among the resistant/immune cultivars, cyst formation was not observed on eggplant, tomato, lettuce, perilla, carrot, celery, watermelon, oriental melon, cucumber, pumpkin, chives, onion, welsh onion, balloon flower roots, deodeok (Codonopsis lanceolata), Jandae (Adenophora triphylla), and bean. Therefore, these plants are regarded as immune to the cyst nematode. However, many crops belonging to Solanaceae, Asteraceae, Chenopodiaceae, and Poaceae families showed moderate susceptibility or immunity, depending on the crop or cultivar. This study provides a basis for alternative crop recommendations for sugar beet nematode cyst-infected farms in Chinese cabbage production areas.

• Journal of Microbiology and Biotechnology
• /
• v.26 no.2
• /
• pp.385-393
• /
• 2016

Pseudomonas syringae pv. actinidiae causes bacterial canker disease in kiwifruit. Owing to the prohibition of agricultural antibiotic use in major kiwifruit-cultivating countries, alternative methods need to be developed to manage this disease. Bacteriophages are viruses that specifically infect target bacteria and have recently been reconsidered as potential biological control agents for bacterial pathogens owing to their specificity in terms of host range. In this study, we isolated bacteriophages against P. syringae pv. actinidiae from soils collected from kiwifruit orchards in Korea and selected seven bacteriophages for further characterization based on restriction enzyme digestion patterns of genomic DNA. Among the studied bacteriophages, two belong to the Myoviridae family and three belong to the Podoviridae family, based on morphology observed by transmission electron microscopy. The host range of the selected bacteriophages was confirmed using 18 strains of P. syringae pv. actinidiae, including the Psa2 and Psa3 groups, and some were also effective against other P. syringae pathovars. Lytic activity of the selected bacteriophages was sustained in vitro until 80 h, and their activity remained stable up to 50℃, at pH 11, and under UV-B light. These results indicate that the isolated bacteriophages are specific to P. syringae species and are resistant to various environmental factors, implying their potential use in control of bacterial canker disease in kiwifruits.

• The Bulletin of The Korean Astronomical Society
• /
• v.38 no.1
• /
• pp.30.2-30.2
• /
• 2013

The mean color of globular cluster (GCs) systems in early-type galaxies (ETGs) is, in general, bluer than the integrated color of field stars in their host galaxies. Recently, Goudfrooij & Kruijssen (2013) reported that even red GCs in the ETGs show bluer colors than their host field stars and suggested the different initial mass function (IMF) for red GCs and field stars to explain the observed offset in color. Here we suggest an alternative scenario that explains the observed color offsets between red GCs in ETGs and the field stars in the parent galaxies without invoking to the variation of the IMF. We find that the inclusion of second-generation (SG) helium-enhanced populations in the model fully explains the observed color offset between red GCs and field stars in the host galaxies. We have also tested the effect of the IMF slope on our models, but the effect is relatively small compared to the effect of the SG population. Our new model suggests that, in order to explain far-UV strong metal-rich GCs in M87 and the observed color offset between metal-rich GCs and the field stars in ETGs simultaneously, the inclusion of the SG populations with enhanced helium abundance is a more natural solution than the model that only adopted variations in the IMF.

• Journal of Microbiology and Biotechnology
• /
• v.34 no.4
• /
• pp.804-811
• /
• 2024

Foamy viruses (FVs) are generally recognized as non-pathogenic, often causing asymptomatic or mild symptoms in infections. Leveraging these unique characteristics, FV vectors hold significant promise for applications in gene therapy. This study introduces a novel platform technology using a pseudo-virus with single-round infectivity. In contrast to previous vector approaches, we developed a technique employing only two vectors, pcHFV lacking Env and pCMV-Env, to introduce the desired genes into target cells. Our investigation demonstrated the efficacy of the prototype foamy virus (PFV) dual-vector system in producing viruses and delivering transgenes into host cells. To optimize viral production, we incorporated the codon-optimized Env (optEnv) gene in pCMV-Env and the Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE) at the 3' end of the transgene in the transfer vector. Consequently, the use of optEnv led to a significant enhancement in transgene expression in host cells. Additionally, the WPRE exhibited an enhancing effect. Furthermore, the introduced EGFP transgene was present in host cells for a month. In an effort to expand transgene capacity, we further streamlined the viral vector, anticipating the delivery of approximately 4.3 kbp of genes through our PFV dual-vector system. This study underscores the potential of PFVs as an alternative to lentiviruses or other retroviruses in the realm of gene therapy.