• The Journal of the Korean Society for Microbiology
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• v.21 no.1
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• pp.33-45
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• 1986

On hundred and forty stains of shigella cultures isolated from the twelve hygiene laboratories of cities and provincial general hospital laboratories in 1983 were tested for their resistance to thirteen antimicrobial drugs and their R-plasmid transfer. Antimicrobial drugs were used amikacin, ampicillin, cephalothin, chloramphenicol, gentamicin, kanamycin, nalidixic acid, rifampicin, streptamycin, tetracycline, tobramycin, cefoperazone and piperacillin. All strains were resistant to one or more of thirteen antimicrobial drugs but 94.3% were susceptible to amikacin, gentamicin and tobramycin of total isolated. The most strains commonly found resistance was to chloramphenicol (94%) followed by streptamycin (93%), tetracyline (92%) piperacillin (90%) ampicillin (83%), cefoperazone (42%), nalidixic acid (14%), cephalothin (17%), rifampicin (22%) and kanamycin (6%), sixty percent of strains among 140 were resistance to ampicillin, chloramphenicol, streptomycin, tetracycline at the same time. The transfer of drug resistance by conjugation was tested and ninety four strains (94.3%) were resistant to one or more drugs were found to transfer their drug resistance of E. coli. percentage of transfer frequency by conjugation was one strains (54%), the transfer frequency of drug resistance varied by donor strains and recipients, but not by selecting drugs. Resistance to nalidixic acid was not transferred by conjugation to recipients. Percentage of plasmid curing after the treatment of acriflavine, acridine orange was about 8%. Among strains cured two strains were tested compare original strains with them in biochemical properties in arginine dihydrolase and arabinose fermentation reaction. It was found to growth curves of No.2 shigella flexneri, serotype 1b, and its derivatives cured with acriflavine in $M{\ddot{u}}ller$ Hinton broth medium (pH 7.4, $38^{\circ}C$) by temperature Gradient Biophoto Recorder TN-1120 (Tokyo, Japan).

• Korean Journal of Microbiology
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• v.28 no.3
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• pp.219-228
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• 1990

A kanamycin resistance($Km^r$) gene was studied for its transfer in natural freshwater environments by using the natural bacterial isolate(M1) of DK1 and the DKC601 strain, $Km^r$ plasmid of which was genetically engineered from the NI strain. The transfer frequency ofthe $Km^r$ gene and rearrangement of the $Km^r$ plasmid were compared between the gnetically engineered microorganism(GEM) and the NI parental strain by conjugation with the same recipient strain. The transfer frequency of the $Km^r$ gene was about $9.1\times 10^{-12}-1.8\times 10^{-11}$ in both the GEM and NI strains at 5 to $10^{\circ}C$, but the frequency of the NI was about 10 times higher than that of the GEM at 20 to $30^{\circ}C$. The $Km^r$ plasmid in the transconjugants obtained by conjugation of the NI with the MY1 strain as a ricipient showed alot of rearrangement, but the $Km^r$ plasmid transferred from the GEM was stable without alteration of its size. When the MT2 strain was used as a recipient, however, such a rearrangement of the $Km^r$ plamid was observed in the transconjugants obtained from the GEM as well as the NI strain. In those transconjugants obtained from different mating pairs and water environments, the plasmid were appeared to decrease in their number as the period of conjugation time was prolonged, but only the $Km^r$ plasmid transferred from the GEM kept having its size of 52kb. Therefore, the $Km^r$ gene was transferred at the same rate from the GEM and NI strains in natural freshwater environment, but the gene of the GEM strain was more stable than the NIduring conjugation and the $Km^r$ plasmid was rearranged by changing the recipient strain for conjugation in any water environments.

• Microbiology and Biotechnology Letters
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• v.20 no.4
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• pp.483-490
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• 1992

In order to understand the transfer behavior of a particular gene in water environments, kanamycin resistance ($Km^r$) gene was tracked by Southern hybridization with DNA probe in its conjugal transfer. A $Km^r$ natural bacterial isolate and genetically modified microorganisms (GMMs) constructed from the isolate were used as donor for conjugal transfer of the $Km^r$ gene. The transfer frequencies of the $Km^r$ gene from GMM strains were generally 10 to 100 times higher than those from the natural isolate. The conjugants obtained from GMM strains in LB broth had more plasmids newly appeared, and particularly the conjugants in A Wand FW waters revealed more rearrangement in their plasmids as a function of conjugation time. When plasmids of the conjugants obtained in LB broth were Southern hybridized with DNA probe of the $Km^r$ gene, the $Km^r$ plasmids in the conjugants were detected at the same position of the plasmids in donor cells, in spite of the fact that the plasmids were highly rearranged in conjugant cells. But the $Km^r$ plasmids in the donor of DKI and DKC601, and DKC600 were not identified in the conjugants obtained after 50 h conjugation in AW and after 30 h in AW, respectively. The size of the $Km^r$ plasmids showing hybridization signal were a little changed in the conjugants obtained in A Wand FW waters. Therefore, the method of Southern hybridization with DNA probe was proved to be very specific and useful for tracking of particular genes in water environments.

• Journal of Microbiology and Biotechnology
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• v.25 no.10
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• pp.1702-1708
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• 2015

We have developed a new shuttle plasmid, designated as pLK1-MCS that can replicate in both Clostridium acetobutylicum and Escherichia coli, by combining the pUB110 and pUC19 plasmids. Plasmid pLK1-MCS replicated more stably than previously reported plasmids containing either the pIM13 or the pAMβ1 replicon in the absence of antibiotic selective pressure. The transfer frequency of pLK1-MCS into C. acetobutylicum was similar to the transfer frequency of other shuttle plasmids. We complemented C. acetobutylicum ML1 (that does not produce solvents such as acetone, butanol, and ethanol owing to loss of the megaplasmid pSOL1 harboring the adhE1-ctfAB-adc operon) by introducing pLK1-MCS carrying the adhE1-ctfAB-adc operon into C. acetobutylicum ML1. The transformed cells were able to resume anaerobic solvent production, indicating that the new shuttle plasmid has the potential for practical use in microbial biotechnology.

• Microbiology and Biotechnology Letters
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• v.23 no.2
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• pp.164-169
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• 1995

An endogenous cryptic plasmid, pBL1, which has been used to construct plasmid vectors for coryneform bacteria producing amino acids, was eliminated from Brevibacterium lactofermentum. The pBL1 was partially digested with Sau3AI and the resulting DNA fragments were subcloned into a suicide vector pEM1 which contains a kanamycin-resistant (km$^{r}$) gene. KM$^{r}$ B. lactofermentum transconjugants were obtained by conjugal transfer of the pEM1 derivatives containing pBL1 DNA fragments from Escherichia coli into B. lactofermentum. A km$^{r}$ transconjugant was analyzed to contain a plasmid pEB14, which occurred in vivo by homologous recombination between pBL1 and the conjugal-transferred plasmid. The pEB14 including the pEM1-derived km$^{r}$ gene was found to be lost concomitantly with km$^{r}$ phenotype, resulting in the construction of a pBL1-free strain of B lactofermentum. Based on transformation efficiencies and plasmid stability, the resultant pBL1- free strain is more useful than wild strain as a host cell for genetic manipulation. It could be concluded that foreign plasmid DNAs are efficiently isolated and analyzed from the pBL1-free strain because of the absence of endogenous pBL1 plasmid.

• Journal of Microbiology and Biotechnology
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• v.17 no.1
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• pp.162-167
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• 2007

A genetic transfer system for introducing foreign genes to biomining microorganisms is urgently needed. Thus, a conjugative gene transfer system was investigated for a moderately thermophilic, extremely acidophilic biomining bacterium, Acidithiobacillus caldus MTH-04. The broad-hostrange IncP plasmids RP4 and R68.45 were transferred directly into A. caldus MTH-04 from Escherichia coli by conjugation at relatively high frequencies. Additionally the broad-hostrange IncQ plasmids pJRD215, pVLT33, and pVLT35 were also transferred into A. caldus MTH-04 with the help of plasmid RP4 or strains with plasmid RP4 integrated into their chromosome, such as E. coli SM10. The $Km^r\;and\;Sm^r$ selectable markers from these plasmids were successfully expressed in A. caldus MTH-04. Futhermore, the IncP and IncQ plasmids were transferred back into E. coli cells from A. caldus MTH-04, thereby confirming the initial transfer of these plasmids from E. coli to A. caldus MTH-04. All the IncP and IncQ plasmids studied were stable in A. caldus MTH-04. Consequently, this development of a conjugational system for A. caldus MTH-04 will greatly facilitate its genetic study.

• Annual Conference on Human and Language Technology
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• 2003.10d
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• pp.293-299
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• 2003

인간 게놈 프로젝트가 완료됨에 다라 생체서열과 언어 사이의 대응 관계가 부각되고 있다. 본고에서는 Lewis Carroll의 언어 유희 사례를 컴퓨터생물학의 측면에서 재조명하고, Carroll이 제시한 문제 중에서 간단한 anagram 문제의 해결을 다루고자 한다. 우선 DNA 컴퓨팅의 방법론을 적용한 DNAgram의 개념을 확장하여 plasmid-DNAgram의 개념을 새롭게 도입하였다. 이 개념을 형광단백질에 대한 DNAgram의 개념을 확장하여 plasmid-DNAgram의 개념을 새롭게 도입하였다. 이 개념을 형광단백질에 대한 FRET(fluorescent resonance energy transfer)분석기법의 응용 사례인 cameleon 형광단백질에 대한 FRET 분석기법에 적용함으로써 anagram 문제의 어휘론적, 구문론적, 의미론적, 화용론적 측면에 대응하는 바이오분자 컴퓨팅 방법론을 제안하였다.

• Journal of Sericultural and Entomological Science
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• v.40 no.1
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• pp.52-59
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• 1998

For the construction of plasmid and bmNPV sarrying the FRT recognition site for the FLP recombinases, we synthesized the wild type FRT dligonucleotides. The target FRT sequences consist of three 13bp repeated DNA sequences; two repeats in a direct orientation and one inverted relative to the other two. In addition, there is an 8bp spacer region between the repeats which determune the orientation of the FRT recombination site. In order to place the FRT site both in target BmNPV genome and the transfer vector, we constructed a plasmid, FRT site both in the target BmNPv genome and the transfer vector, we constructed a plasmid, pFRT$\beta$-gal, carrying the FRT sites within the cloning sites of pSV vector and a recombinant BmNPV, vFRTPH, carrying the FRT sites at a downstream of polyhedrin promotor, respectively. In order to test the functionality of the FLP/FRT site-specific recombination system, vFRTPH, pFRT$\beta$-gal and pHsFLP DNA were co-transfected into BmN-4 cells. The resulting recombinant virus was designated a vFRT$\beta$2-gal. From construction analysis of the vFRT$\beta$2-gal with PCR technique it was concluded that the entire pFRT$\beta$-gal plasmid with $\beta$-galactosidase gene and origines of replication flanked by two functional hybrid FRT sequences. The efficiency of recombination was 8.7%, which was higher than that(2.2%) of recombination between a conventional transfer vector and the wild type BmNPV.

• Journal of Microbiology and Biotechnology
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• v.15 no.2
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• pp.376-383
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• 2005

Alcaligenes sp. JMP228 carrying 2,4­dichlorophenoxyacetic acid (2,4-D) degradative plasmid pJP4 was inoculated into natural soil, and transfer of the plasmid pJP4 to indigenous soil bacteria was investigated with and without 2,4-D amendment. Plasmid pJP4 transfer was enhanced in the soils treated with 2,4-D, compared to the soils not amended with 2,4-D. Several different transconjugants were isolated from the soils treated with 2,4-D, while no indigenous transconjugants were obtained from the unamended soils. Inoculation of the soils with both the donor Alcaligenes sp. JMP228/pJP4 and a recipient Burkholderia cepacia DBO 1 produced less diverse transconjugants than the soils inoculated with the donor alone. Repetitive extragenic palindromic-polymerase chain reaction (REP-PCR) analysis of the transconjugants exhibited seven distinct genomic DNA fingerprints. Analysis of 16S rDNA sequences indicated that the transconjugants were related to members of the genera Burkholderia and Pandoraea. Denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S rRNA genes revealed that inoculation of the donor caused clear changes in the bacterial community structure of the 2,4-D­amended soils. The new 16S rRNA gene bands in the DGGE profile corresponded with the 16S rRNA genes of 2,4-D­degrading transconjugants isolated from the soil. The results indicate that introduction of the 2,4-D degradative plasmid as Alcaligenes sp. JMP228/pJP4 has a substantial impact on the bacterial community structure in the 2,4-D-amended soil.

• KSBB Journal
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• v.19 no.5
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• pp.341-347
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• 2004

Several gene delivery therapies are being developed for treatment of serum protein deficiency. EPO is one of the most promising therapeutic agent for this treatment which is currently being investigated in depth. This study has the ultimate purpose of improving the gene delivery system for an increase of red blood cell production. A plasmid DNA was constructed smaller than other plasmids for an increase in penetration into animal cells, and two genes were cloned into each vector as a co-delivery system to express erythropoietin, and interluekin-3 or thrombopoietin, which can act on erythroid cell, thus activating hematopoiesis synergically. This co-delivery system has an advantage of decreasing the labour required for industrial production of DNA vaccine. A new plasmid vector, pVAC, in size 2.9 kb, was constructed with the essential parts from PUC 19 and pSectagB, which is much smaller than other plasmid vector and is the size of 2.9 kb. Co-delivery system was constituted by cloning human erythropoietin with each of human interluekin-3 gene or human thrombopoietin gene into both pVAC and pSectagB. As a result, the transfection efficiency of pVAC was higer than that of pSectagB in vitro, and hematocrit level of the mice injected with pVAC is higher than that of other mice. And co-delivery system, made of several plasmid DNAs, was expressed in vitro.