• Journal of Microbiology and Biotechnology
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• v.6 no.4
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• pp.292-294
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• 1996

To investigate the relationship between two tetracycline resistance ($Tc^r$) plasmids, the 24.82-kb pKHl and the 4.44-kb pKH6, of Staphylococcus aureus isolated in Korea, cloning of the 4570-bp HindIII fragment into pBluescript II $KS^+$ after partial digestion of the $Tc^r$ plasmid pKHl with HindIII and sequence determination of that fragment were carried out. Analysis of the sequences revealed that the 4570-bp HindIII fragment contained a 4011-bp fragment of the small $Tc^r$ plasmid pKH6 flanked by the partial sequences of IS431mec. It was concluded from the above result that the pKHl was produced by integration of the partial sequence of the pKH6 into another plasmid via IS431mec.

• Korean Journal of Microbiology
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• v.26 no.4
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• pp.298-304
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• 1988

Antibiotic resistance plasmids (RP4, pMG1, R679 and R91-5) were used as primary selective markers to detect the fusants in Pseudomonas spp. By using plasmid marker, clones containing both plasmids of parental strains were obtained as fusants by direct selection. The spheroplast fusion was occured not only between strains of interspecies, but also between strains of intergenus such as Pseudomonas and E. coli. The frequencies of fusant formation were variable from $2.8\times 10^{-1}$ to $6.0\times 10^{-4}$ -4/. In addition, chromosomal recombinants were formed among the clones with both parental plasmids in frequencies of 0.44-1.3%. The fusant formation frequency between interspecies of intraspecies was not different markedly, but stability of plasmids in fusants correlated with the phylogenic smilarity of the parental strains.

• Korean Journal of Microbiology
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• v.22 no.4
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• pp.249-255
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• 1984

To develop the host-vector system for industrial Coryneform bacteria that seemed to be the most suitable microorganisms for molecular breeding of genes involved in the production of amion acids, nucleotides, and other products of industrial interest, broad host range E. coli plasmid R 1162 DNA was transformed into Brevibacterium ammoniagenes and the plasmids pKU6 isolated from a transformant was physically characterized. All other plasmids from the transformed cells except pKU6 exsisted as multimeric forms in Brevibacterium ammoniagenes. The plasmid DNA was retransformed into Corynebacterium glutamicum with a high frequency ($1.32{\times}10^{-1}$ per cell) and maintained stably both in Brevibacterium ammoniagenes and Corynebacterium glutamicum after 100 generations of cultures with 25-30 copy number per cell. The size of both plasmid pKU6 and plasmid R1162 were the same and restriction maps by EcoR I, Ava I, Pst I, Pvu II and Hinc II were also similar.

• YAKHAK HOEJI
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• v.34 no.1
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• pp.54-63
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• 1990

These studies were made to assess the present stage of resistance of Shigella species to antibiotics and to find characteristics of R-plasmid of these bacteria. From 1986 to 1988, 125 strains of Shigella species were isolated from patients specimens collected in Chung Cheong-do Hospital, Kyunghee Medical Center, city an provincial health & environmental institutes. These specimens were tested for resistance to 12 kinds of antimicrobial agents by agar dilution method. Using Muller-Hinton agar for the assay of drug resistance and Trypticane Soy Broth as propagating medium for conjugation. All the strains (100%) were resistant to one or more antibiotics. Drug resistance patterns of isolated strains were found as the highest resistance to ampicillin (98%) in 1986, to tetracycline (98%) in 1987, to tetracycline (100%) in 1988, all strains were sensitive to gentamicin, amikacin, tobramycin. Chronologically, resistance decreased gradually as it was shown in relation to kanamycin, rifampicin in 1986, 1987 and 1988, (4%, 2%) (4%, 2%) (0%, 0%) respectively. But, resistance was increased year by year as it was shown in relation to tetracycline, nalidixic acid, streptomycin in 1986, 1987, 1988 (89%, 19%, 45%) (98%, 46%, 71%) and (100%, 58%, 88%). The resistance in correlation to more than 5 drugs, which was 13 strains among 47 strains in 1986, 38 strains among 87 strains in 1987, 23 strains among 26 strains in 1988, was increased gradually. In the transfer test of drug resistance by conjugation methods, the rate which was 3 strains (50%) in 1986, 8 strains (62%) in 1987, 3 strains (100%) in 1988, was increased gradually. When the donor strains were conjugated with the recipient strains, the conjugation rate was high in the multiple resistant strains. The relationships of transferring patterns of drug resistance and molecular weight of R-plasmid were variable. However, only a plasmid which has more than 35 Mgd was transferred.

• Microbiology and Biotechnology Letters
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• v.14 no.3
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• pp.245-250
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• 1986

Antibiotic resistant bacteria isolated from the river water in Cheongju City were studied on the transfer of their R-plasmids in water environment. The R-plasmids were transferred by conjugation between the isolates ai the frequencies of 1.1$\times$10$^{-6}$ to 1.2$\times$10$^{-7}$ under laboratory condition and 1.2$\times$10$^{-7}$ to 1.0$\times$10$^{-9}$ in natural environment. The R-plasmids isolated from those bacteria were also transferred into the recipient cells of E. coli HB101 at the frequencies of 1.7-6.7$\times$10$^{-6}$. The AP$^{r}$Cm$^{r}$Tc$^{r}$-plasmid of isolate T-44 which were transformation by conjugation and transformation was determined to be 9.01 kilobses in molecular size. When the AP$^{r}$Cm$^{r}$Tc$^{r}$-plasmid DNA was treated with restriction endonuclease, the plasmid appeared to have one restriction site for EroRI and BamHI, respectively, and three sites for Pst I endonuclease.

• 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.

• Korean Journal of Microbiology
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• v.30 no.2
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• pp.88-95
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• 1992

The genetically engineered microorganisms (GEMS) could be released accidentally or ii)rexperimental purposes, as the genetic engineering, technique ha:. become very popular inany laboratories of biological sciences. But there have been littlt: informations on transkrbehavior of the genetically ~nodified genes in the natural en\ironmentx. In this stutly.antibiotic resistant bacteria were isolated from nat.ural waters. and then GEM strains wereconstructed i'rom the natural isolate (NI) by ~noclification oi' the Km' plasmitl. Thetransferability of the plasmids in the GEM and NI strains were examinetl by con-jugationin Luria-Bertani broth :it 30$^{\circ}$C. Also the cff'ccts 01' mating strain and pH on their transferfrequency and rearrangement of the plasmids in tl-~ec o~ijugantsM ere comp:irati\ely stuclictl.I'hc transkr frequency of Km' plasmid in donor of GEM and N1 strains wah similar a.;about 10 ' when co~ljugation was conducted wit11 M'I'I strain is recipient at pH 7. butthat of 1)KCOOI was lowered to 1.2X 10 '. And when the lab. stlain was uhccl as recipient.the transfer tendency of the plasmid was about same in both (;EM and NI strains usedas donor. All thc tionor 5trains. except for I)KC601. showecl the Ilighcs~ frequency of about10 ' at pH 7 and the frequcncics were lowered at both pH 5 and 9. Hut the mocliliedKm' plasmid in the cloned strain of DKC601 was transferred hy very low frequency of10 "at pH 5 ant1 7 comparing to other GEM strains. especiall! any co~~.jugantws ere notobtained at pH 4 and 9 even after conjugation for 6 hours. Rearrangement of the plasmidstranskrred into the lab. strain was not found in the conjugants. I\ulcornerut a lot of rearrangclncntwas ohservecl nlhen they were transferred into the NI strain. Such a rearrangement wasmore severe when donor was GEM strain rather then NI strain Hut such ;r phenomenonwas less affected by p!-l values.r phenomenon was less affected by p!-l values.

• Microbiology and Biotechnology Letters
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• v.11 no.1
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• pp.75-79
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• 1983

To exploit a suitable vector and recipient strain for molecular cloning in Bacillus subtilis, oxytetracycline-resistant plasmic DNA has been prepared from Streptomyces rimosus by phenol-buffer extraction of lysozyme-lysed cells and introduced into B. subtilis KPM 60 [St $r^{R}$-mutant of RM 125 (leu A8, arg 15, hsm $M^{-10}$ , hsr $M^{-10}$ )] by transformation. Oxitetracycline-resistant plasmid was well transferred into B. subtilis KPM 60 with average frequency of 10$^{-4}$ per $\mu\textrm{g}$ of DNA. The highest frequency of plasmid transformation was obtained after 3 hours incubation of recipient cells in the growth medium and 30 to 60 minutes incubation in the competence medium, and then 20 minutes contact of DNA and host cells. Optimum pH for competence was 7.5, and optimum temperature for transformation was 2$0^{\circ}C$.>.

• Korean Journal of Microbiology
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• v.25 no.4
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• pp.282-289
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• 1987

The characteristics of the plasmid pKU10 isolated from Pseudomonas putida KU816 were investigated and its restriction map was constructed. The pKU10 plasmid was a small R plasmid carrying genes for resistance to ampicillin, tetracyclin, and chloramphenicol, and cured by treatment with mitomycin C. The molecular size of pKU10 was estimated to be 9.4Kb. Pseudomonas strains and E. coli cells could be transformed for antibiotic resistance characters specified by pKU10 plasmid DNA. By incompatibility test with other plasmids, pKU10 is grouped into IncP-1. EcoRI, XhoI, SalI, BglII, and SmaI cleaved pKU10 once, while PstI cleaved at two sites, and HindIII cleaved at six sites. The restriction map was constructed by partial and complete digestion of the purified plasmid DNA with single, double, or triple restriction enzymes. Thus, pKU10 is expected to be used for a cloning vector in Pseudomonas cells.

• Korean Journal of Microbiology
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• v.40 no.2
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• pp.172-177
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• 2004

Photosynthetic bacteria, Rhodospirillum rubrum, have been reported to change their metabolic patterns depend-ing on the light condition. The genetic approach for such a metabolic change is one of main subject in pho-tosynthetic bacteria. It has been reported that the extrachromosomal plasmid might be related to this metabolic regulation. In this study, we have determined the partial sequences of R. rubrum plasmid pKYl with HindIII fragments and the predicted pKYl ORFs and physical map. We found the 8 putative proteins related to the genetic recombination of bacterium, which is reported to the alternative gene expression. Our results suggest that the genes located in pKYl are possibly involved in the metabolic switch according to the photocondition.