• The Korean Journal of Food And Nutrition
• /
• v.9 no.4
• /
• pp.459-465
• /
• 1996

Bacillus thuringiensis serovar. darmstadiensis produced bipyramidal endo-toxin. The toxin protein was purified by Renografin-76 step gradient centrifugation and investigated by electron microscope. Analysis of total plasmid DNA patterns showed that four different size of plasmids existed in wild type B. thuringiensis serovar. darmstadiensis. Total plasmids DNA was isolated and transformed into pst I site of pBR322 cloning vector. Ten clones containing crystal toxin gene were forst screened colony hybridization by using PUYBT 9044 probe ontained B. thuringiensis kurskaki HD 1 toxin gene. Cloned-DNA was digested with EcoR1 and HindIII and transformed to pIBI30 sequencing vector. Finally, 2.6kb and 3.6kb size fragments contatined toxin-gene were cloned with restriction analysis.

• The Journal of the Korean Society for Microbiology
• /
• v.22 no.3
• /
• pp.241-250
• /
• 1987

A total 64 strains of Escherichia coli including 38 strains of urinary tract infection and 26 strains from other clincal sources were studied for several properties related to the virulence markers of organisms. Urinary isolates(76.3%) showed higher frequency of mannose resistant hemagglutination(MRHA) wi th human erythrocytes(A type, $Rh^+$) than the strains of control group isolated from other sources(34.6%). Seventeen strains(44.4%) of urinary isolates and 2 strains(7.7%) of control group showed hemolysis on blood agar plate. There was no significant difference in MIC's of 23 drugs between both groups of urinary isolates and control group. But they showed high frequency of resistance to ampicillin, carbenicillin, piperacillin, kanamycin, and trimethoprim, but were very susceptible to cefotaxime, moxalactam, ceftizidime, imipenem, and norfloxacine. Fourteen strains(36.8%) of urinary isolates and 10 strains(38.5%) of control group showed conjugally transferable resistance conferred to R plasmids. The urinary isolates carried one or more to 6(mean 3.4) plasmids of approximate molecular weight ranged 3.1 to 94 megadalton(Mdal) and strains of control group carried 2 to 5(mean 3.8) plasm ids of size ranged 3.6 to 130 Mdal. The size of conjugally transferable R plasmid identified with transconjugants ranged 32 to 130 Mdal.

• Korean Journal of Microbiology
• /
• v.21 no.2
• /
• pp.95-102
• /
• 1983

The penicillin G acylase(pga) gene was cloned in the vector plasmid pKM $300(Ar^r,\;Tc^r,\;6.33kb)$ for the study of the structure and expression of the pga gene. This recombinant plasmid pPAKS-1 DNA(24.5 Kb) was cleaved into 2 fragments by restriction enzyme Eco R1.1fragment by BamH1, 4fragments by Hind III, and 2 fragments by Pst I. The pga gene was located on the Eco R1.Hind III-C fragement of pPAKS-1. The recombinant plasmids pPAKS-1 and pPAKS-2, in which the Hind III-B and Hind III-D fragments pPAKS-1 are deleted, are characterized. The results are summarized as follows : 1. Doubling times of bacterial strain bearing pPAKS-1 and pPAKS-2 are 90 and 60 minutes, respectively. 2. pPAKS-1 and pPAKS-2 are present at about 16-32 and 70 copies per cell, respectively, are 0.66 and 5.5 units, respectively, which represent 2-fold and 20-fold higher enzyme 4. pPAKS-1 is very unstable, but pPAKS-2 is stable.

• Korean Journal of Microbiology
• /
• v.31 no.1
• /
• pp.44-47
• /
• 1993

A replication initiation gene was identified and its nucleotide sequence has been determined from a 3.8 kb, chloramphenicol acethyltransferase conferring R-plasmid pSBK203 of Staphylococcus aures. Location of the replication related region of pSBK 203 was determined by interuption with pUC 119 at XBaI and MspI sites which resulted in inactivation of replication in Bacilius subtilis. Base sequence of this region revealed on open reading frame of 942 base pairs, which encoded a 314 amino acid protein. Base sequence homology with other rep of pT181 family plasmids such as pT181, pC221, pC223, pS194, pU112, and pCW7 was ranged from 78% to 97% and the predicted amino acid sequence homology was from 72% to 95%.

• The Journal of the Korean Society for Microbiology
• /
• v.14 no.1
• /
• pp.49-61
• /
• 1979

Two hundred and ninety-five strains of Peudomonas aeruginosa isolated from clinical sources were tested for drug resistance and demonstration of R plasmids by intraspecies conjugation system. Sixty strains were found highly resistant to two or more of drugs. The rate of resistant strains were 38.9% to kanamycin(km), 33.2% to streptomydn(sm), 22.7% to sulfisomidine(Sa), 14.2% to chloramphenicol(Cp), 13.8% to tetracycline(Tc), 3.0% to carbenicillin(Cb), and to gentamicin(Gm), respectively. But no strains was resistant to nalidixic acid and colistine. They were resistant to per milliliter to more than $400{\mu}g$ per ml. of Tc, $800{\mu}g$ per ml of Cp and of Sm, $6,400{\mu}g$ per ml. of Sa, $200{\mu}g$ per ml. of Cb, $100{\mu}g$ per ml. of Gm, and $25{\mu}g$ per ml. of colistine. Forty-three strains of Pseudomonas aeruginosa could be transferred their resistance to Pseudomonas aeruginosa 2-70, 1005 rifampin resistant FP-auxotrophic mutant. Of sixty multiple resistant strains, forty-three(71.6%) demonstrated R plasmids; nineteen carried resistance to(Tc Cp Sm Sa), six to(Tc Cp Sm), three to(Tc Cp Sa), and Cp, five to(Tc Sm Sa), two to(Tc Sa), (Cp Sm) and Tc, and one to(Cp Sm Sa). Degree of resistance of recipients recieving R plasmids from donors were almost the same level of resistance as the donor in regardless of mating temperature at $25^{\circ}C$ and $37^{\circ}C$. Resistance to Tc, Sm, and Sa were transferred to a very few of recipient cells at five minutes after mating with donor and recipient cells but resistance to Cp were transferred to the majority of recipient cells. The transfer frequency of Tc, Cp, Sm, and Sa resistance from donors to recipients were from $1.0^{-1.4}\;to\;1.0^{-3.5}$ at $25^{\circ}C$ for 18 hours of incubation and were from $1.0^{-1.5}\;to\;1.0^{-3.5}$ at $37^{\circ}C$ for 18 hours of incubation.

• Asian-Australasian Journal of Animal Sciences
• /
• v.18 no.9
• /
• pp.1336-1342
• /
• 2005

The present research work was conducted to evaluate the beneficial effects as well as the safety aspects of lactobacilli as probiotic. Lactobacilli were isolated from poultry faecal samples, feed samples and from some known preparations procured from poultry feed manufacturers. L. acidophilus and L. sporogenes were tested for the antibacterial activity against four poultry pathogens viz. Escherichia coli, Salmonella spp., Proteus spp. and Pseudomonas aeruginosa. Cell free supernatant (CFS) of L. acidophilus exhibited significantly higher antibacterial activity against Salmonella spp. at original pH (4.50${\pm}$0.02). At the adjusted pH (6.50${\pm}$0.02) significantly higher antibacterial activity was recorded against indicator organism except for P. aeruginosa. Likewise, L. sporogenes exhibited similar antibacterial activity at original as well as adjusted pH except for E. coli. Antibacterial activity against E. coli was significantly higher at adjusted pH than at original pH of CFS. The competitive exclusion of E. coli by lactobacilli over the intestinal epithelial cells (IEC) was checked. L. acidophilus strain I, which was of poultry origin, exhibited maximum attachment over IEC as compared to other three strains of non-poultry origin viz. L. acidophilus strain II, L. sporogenes strain I and II. Overall, L. acidophilus exhibited higher competitive exclusion as compared to L. sporogenes. All the lactobacilli of poultry origin were most sensitive to penicillin G, amoxycillin, ampicillin and chloramphenicol, least sensitive to sulphamethizole, ciprofloxacin, neomycin, norfloxacin and pefloxacin and resistant to metronidazole and nalidixic acid. The isolates from probiotic preparations were most sensitive to ampicillin, amoxycillin and tetracycline, least sensitive to sulphamethizole, norfloxacin, neomycin and ceftriazone and resistant to nalidixic acid and metronidazole. Eight of the multiple drug resistant lactobacilli isolates were studied for the presence of plasmids. Plasmids could be extracted from six isolates of lactobacilli. These plasmids could be responsible for bacteriocin production or for antibiotic resistance of the strains. The lactobacilli need further studies regarding their safety for use in the probiotic preparations.

• Journal of Microbiology and Biotechnology
• /
• v.27 no.9
• /
• pp.1711-1715
• /
• 2017

In this study, we characterized the $bla_{NDM-5}$-bearing plasmid in a Klebsiella pneumoniae isolate that had lost the plasmid during serial passage. We determined the complete sequences of the plasmid pCC1410-2, which was extracted from a K. pneumoniae ST709 isolate collected at a Korean hospital from which two NDM-5-producing K. pneumoniae isolates were subsequently isolated. As a result, the pCC1410-2 plasmid had a backbone structure that was similar to those of two plasmids previously reported from the same hospital, but lacked some antibiotic resistance genes ($bla_{TEM-1}$, rmtB, mphR(A), mrx(A), and mph(A)). A 9-bp repeating unit encoding three amino acids (Gln-Gln-Pro) was inserted in TraD in pCC1410-2. Thus, the pCC1410-2 plasmid might be transferred from the previously identified carbapenem-resistant K. pneumoniae, but some delections and inversions might have occurred during the process. We compared the transfer frequency and stability of the plasmids. The relative frequency of conjugative transfer and stability in the host were significantly lower in pCC1410-2 than in previously reported $bla_{NDM-5}$-bearing plasmids in Korea. A low transfer frequency and instability in the host may cause underestimation of carbapenemase-producing Enterobacteriaceae in the clinical setting and in surveillance studies.

• Journal of fish pathology
• /
• v.12 no.2
• /
• pp.115-121
• /
• 1999

MIC test of 16 chemotherapeutic agents was performed on 24 isolates of Edwardsiella tarda collected from flounders. They revealed resistance against combinations of ampicillin, amoxicillin, erythromycin, flumequine, doxycycline(DOXY), nalidixic acid, novobiocin, oxolinic acid, oxytetracycline(OTC), thiamphenicol(TP) and sulfonamide. Two strains carried transferable R plasmid encoding Otc Kanamycin Tp and Otc chloramphenicol Doxy Tetracycline Tp, respectively. The R plasmids were not similar each other on the basis of their digestion pattern of restriction endonuclease, suggesting distribution of different transferable R plasmid among E. tarda from flounders.

• 한국생물공학회:학술대회논문집
• /
• 2005.10a
• /
• pp.687-687
• /
• 2005

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.11 no.2
• /
• pp.116-120
• /
• 2006

We constructed four recombinant plasm ids to enhance the production of clavulanic acid (CA) in Streptomyces clavuligerus NRRL3585: (1) pIBRHL1, which includes ccaR, a pathway-specific regulatory gene involved in cephamycin C and CA biosynthesis; (2) pIBRHL2, containing claR, again a regulatory gene, which controls the late steps of CA biosynthesis; (3) pGIBR containing afsR-p, a global regulatory gene from Streptomyces peucetius; and (4) pKS, which harbors all of the genes (ccaR/ claR/ afsR-p). The plasmids were expressed in S. clavuligerus NRRL3585 along with the $ermE^*$ promoter. All of them enhanced the production of CA; 2.5-fold overproduction for pIBRHL1, 1.5-fold for pIBRHL2, 1.6-fold for pGIBR, and 1.5-fold for pKS compared to the wild type.