• Journal of Microbiology and Biotechnology
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• v.10 no.3
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• pp.312-320
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• 2000

Abstract The full sequence of the plasmid pKJ36, which was derived from Bifidobacterium longum KJ, was determined and analyzed to construct shuttle vectors between E. coli and Bifidobacterium. The plasmid pKJ36 was composed of 3,625 base pairs with a 65.1% G+C content. The structural organization of pKJ36 was highly similar to that of pKJ50, and the three major ORFs on pKJ36 showed high amino acid sequence homologies with those of pKJ50. The putative proteins coded by these three ORFs were designated as RepB (32.0 kDa, pI=9.25), MembB (29.0 kDa, pI=12.25), and MobB (39.0 kDa, pI=IO.66), respectively. The amino acid sequence of RepB showed a 57% identity and 70% similarity with that of the RepA protein of pKJ50. Upstream of the repB gene, the so-called iteron sequence was directly repeated four-and-ahalf times and a conserved dnaA box was identified. An amino acid sequence comparison between the MobB and MobA of pKJ50 revealed a 48% identity and 61 % similarity. A conserved oriT sequence with an inverted repeat identical to that of pKJ50 was also found upstream of the mobB gene. A hydropathy analysis of MembB revealed four possible transmembrane regions. The expressions of the repB and membB genes were confirmed by RT-PCR. The in vitro translation reaction of pKJ36 showed protein bands with anticipated sizes with respect to each putative gene product. S 1 endonuclease treatment and Southern hybridization suggested that pKJ36 replicates by a rolling circle mechanism via a single-stranded DNA (ssDNA) intermediate. A shuttle vector between E. coli and Bifidobacterium sp. was constructed using the pKJ36, pBR322, and staphylococcal chloramphenicol acetyl transferase (CAT) gene. The successful transformation of the Bifidobacterium strains was shown by Southern hybridization and PCR. The transformation efficiency differed from strain to strain and, depending on the electroporation conditions, with a range between $1.2{\times}10^1-2.6{\times}10^2{\;}cfu/\mu\textrm{g}$ DNA.X> DNA.

• Microbiology and Biotechnology Letters
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• v.33 no.4
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• pp.241-247
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• 2005

A new lactococcal shuttle/expression vector for lactococci, pWgal13T, was constructed using a $\beta$-galactosi-dase gene (lacZ) from Lacfococcus lactis ssp. lactis ATCC 7962 as a screening marker. The pWgal 13T was introduced into Escherichia coli DH5a and L. lactis MG1363, and was easily detected by the formation of blue colonies on a medium containing X-gal without any false transformants. Also, the quantitatively lacZ activity of pWgal13T was measured in L. lactis ssp. cremoris MG1363, and was found to be four times higher than that of L. lactis ssp. lactis ATCC7962 grown on a medium containing glucose, which shows that the lacZ gene of pWgal13T can be used for the efficient screening of L. lactis on general media. The pWgal13T was equipped with a lactococcal replicon of pWV01 from L. lactis Wg2, the new promoter P13C from L. lactis ssp. cremoris LM0230, multiple cloning sites, and a terminator for the expression of a relevant gene. The vee-tor pWgal13T was used for the expression of the EGFP gene in E. coli and L. lactis. These results show that the lactococcal expression/shuttle vector constructed in the present study can be used for the production of foreign proteins in E. coli and L. lactis.

• Journal of the Korean Society for Aeronautical & Space Sciences
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• v.45 no.12
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• pp.1069-1075
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• 2017

With the recent development of space technology, the satellite market, especially the small satellite market, is growing globally. As the satellite market continues to grow, the launch vehicle market is also growing, and demand for low-cost launches is increasing. There are a number of options for low-cost launches, including development of engine that uses low-cost propellants, product and transportation cost savings, but the most effective way to reduce launch costs is to reuse the used launch vehicles. USA's Space Shuttle, a famous rocket as manned spacecraft, could be referred as the start of reusable launch vehicle. However, Space Shuttle had limited reusable parts and it was very expensive even though it is a reusable launch vehicle because of its low efficiency. In recent years, aiming at a real reusable launch vehicle, reusable launch vehicle for commercial purposes have been developed around USA's SpaceX and Blue Origin, and re-landing tests were successfully accomplished. In addition, SpaceX successfully did the re-using of first-stage launch vehicle that had been succeeded in re-landing already. In accordance with this trend, countries such as Europe and India are also concentrating on the study of reusable launch vehicles. Including Blue Origin, companies like Virgin Galactic and XCOR in the United States, are also trying to commercialize the same reusable technology as the private manned space tourism. Confirmation of these technology trends is essential, because the re-use technology could change the landscape of the global launch vehicle market.

• Korean Journal of Microbiology
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• v.52 no.1
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• pp.110-115
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• 2016

A small plasmid (pDK4) from the Antarctic marine organism Pseudoalteromonas sp. PAMC 21150, was purified, sequenced and analyzed. pDK4 was determined to be 3,480 bp in length with a G+C content of 41.64% and contains three open reading frames encoding a replication initiation protein (RepA), a conjugative mobilization protein (Mob) and a hypothetical protein. PCR-amplified pDK4 was cloned in high-copy pUC19 to yield the fusion vector pDOC153. The chloramphenicol resistance gene was inserted into pDOC153 to give an ampicillin and chloramphenicol-resistant, Pseudoalteromonas - Escherichia coli shuttle vector (7,216 bp; pDOC155). The TonB-dependent receptor (chi22718_IV ) and exochitinase (chi22718_III ) genes from Arctic marine P. issachenkonii PAMC 22718 were cloned into pDOC155 to produce pDOC158 and pDOC165, respectively. Both vector derivatives were transferred into plasmid-free Pseudoalteromonas sp. PAMC 22137 by the triparental mating method. PCR experiments showed that the genes were stably maintained both in Pseudoalteromonas sp. PAMC 22137 and E. coli $DH5{\alpha}$ cells, indicating the potential use of pDOC155 as a new gene transfer system into marine Pseudoalteromonas spp.

• Proceedings of the Korean Society of Precision Engineering Conference
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• 2008.11a
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• pp.415-416
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• 2008

• Journal of Power Electronics
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• v.5 no.2
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• pp.166-172
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• 2005

The basic equations of a doubly-fed long-stator linear motor for a shuttle-based railway system are established. They show which degrees of freedom exist for controlling the motor. The ratio of stator and rotor current proves to be an important parameter in determining the design of motors, converters and mechanics.

• Proceedings of the Korean Society of Precision Engineering Conference
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• 2013.05a
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• pp.1111-1112
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• 2013

• Proceedings of the Korean Society of Precision Engineering Conference
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• 2013.05a
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• pp.1359-1360
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• 2013

• Proceedings of the Korean Institute of Navigation and Port Research Conference
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• 2007.12a
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• pp.230-232
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• 2007

• 제어로봇시스템학회:학술대회논문집
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• 1986.10a
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• pp.448-451
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• 1986

An automatic welding system is designed for a part of fork lift. The system is composed of articulated type welding robot, welding positioners, shuttle for robot, system controller and welding equipment. From the application of the system, stable weld quality and production cost saving are achieved. In this paper, the hardware features and control structure of the system are presented.