• Title/Summary/Keyword: pSBk203

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Cloning and Base Sequence Determination of Replication Initiation Gene (rep) Isolated from Staphylococcus aureus DH1 R-plasmid pSBK203 (Staphylococcus aureus DH1에서 분리된 R-plasmid pSBK203의 복제 개시 유전자(rep) 분리 및 염기서열 결정)

  • Park, Seung-Moon;Kwon, Dong-Hyun;Byeon, Woo-Hyeon
    • Korean Journal of Microbiology
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    • v.31 no.1
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    • pp.44-47
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    • 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%.

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Cloning of ori region of R-plasmid pSBK203 and construction of new shuttle-vectors for E. coli & B. subtilis using cloned fragments (R-plasmid pSBK203의 ori 부위 재조합 및 이를 이용한 E.coli와 B.subtilis 간의 Shuttle-Vector 구성)

  • 권동현;석종성;변우현
    • Korean Journal of Microbiology
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    • v.25 no.4
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    • pp.262-273
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    • 1987
  • The replication region of the chloramphenical resistance plasmid pSBK203 of Staphylococcus aureus was cloned using pBR322 and pBD9 as vectors. Cloned replication tegion and chloramphenicol resistance gene were recombined to pBR322. The reconstructed vector behaved as a shuttle vector for E. coli and B. subtilis.

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Replication origin (ori) of R-plasmid pSBK203 Isolated from Staphylococcus aureus DHI (Staphylococcus aureus DH1에서 분리한 R-plasmid pSBK203상의 복제개시 부위 ori에 관한 연구)

  • Min, Kyung-Il;Byeon, Woo-Hyeon
    • Korean Journal of Microbiology
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    • v.32 no.3
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    • pp.186-191
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    • 1994
  • The origin of the leading strand replication (ori) and of lagging strand replication (M-O) of R-plasmid pSBK203 was identified and its base sequence was determined. About 50 bp of ori sequence residues overlapped with the structural gene of rep. Sequence comparison reveals that pSBK-ori shares obvious identities with those of pT181 family and consists of two regions, one is conserved and the other is variable region. Of two palindrome sequence located one after another in upstream region of rep gene, palA' instead of palA which shares sequence homology with diverse family of plasmids such as pOX6, pC194, and pE194 seems to act as a signal for conversion of primarily replicated ssDNA to dsDNA (minus origin (M-O)).

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Nucleotide Sequence and Inducibility Analysis of Chloramphenicol Acetyltransferase Gene from Staphylococcus aureus R-plasmid pSBK203 (Staphylococcus aureus에서 분리된 R-plasmid pSBK203상의 chloramphenicol acetyltransferase 인자의 염기서열 및 유발성 분석)

  • 권동현;변우현
    • Korean Journal of Microbiology
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    • v.27 no.3
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    • pp.194-200
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    • 1989
  • The nucleotide sequence of inducible chloramphenicol acetyl-transferase(CAT) gene isolated from a small plasmid pSBK203 of Staphylococcus aureus was determined. The base sequence shows that structural gene of pSBK203-CAT encodes a protein of 213 amino acids and has a leader region which encodes a short polypeptide of 9 amino-acids in its upstream. vertical bar /sup 35/S vertical bar-Methionine labelled CAT gene product in minicell showed almost same mobility with pC194-CAT of which molecular weight is 24Kdal on polyacrylamide gel electrophoresis. Predicted amino acid sequence of pSBK203-CAT has revealed a high degree of homology with the CATs of pC194 and pC221 than those of cat-86, Tn9 and proteus mirabilis PM13.

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Cloning, Base Sequence Determination and Homology Analysis of Replication Controlling cop Gene of R-plasmid pSBK203 Isolated from Staphylococcus aureus DHI (Staphylococcus aureus DH1에서 분리된 R-plasmid pSBK203의 복제조절 유전자 cop의 Cloning, 염기서열 결정 및 상동성 분석)

  • Park, Seung-Moon;Byeon, Woo-Hyeon
    • Korean Journal of Microbiology
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    • v.32 no.2
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    • pp.115-119
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    • 1994
  • Replication control region of pSBK203, a chloramphenicol acetyltransferase conferring plasmid from Staphylococus aureus was cloned and its nucleotide sequence has been determined. Base sequence homology of this copy control region with those of plasmids belonging to pT181 family was obtained and analyzed. Copy number of four copy mutants derived by addtion or deletion of nucleotides in unique XbaI recognition site in copy control region of pSBK203 was also determined.

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R-plasmids in staphylococcus aureus (Staphylococcus aureus의 항생제 내성 plasmid에 관한 연구)

  • 변우현;김영선;조은희;권동현;이호주;홍순주
    • Korean Journal of Microbiology
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    • v.23 no.4
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    • pp.282-290
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    • 1985
  • Small size antibiotic resistance plasmids having molecular weights less than 10 Mdal were isolated and characterized from ten clinically isolated multiple resistant Staphylococcus aureus. Agarose gel electrophoresis profiles and antibiotic resistance patterns divided these strains into four groups. Strain 2-23-6, the representative strain of a group of five strains conferred two plasmids of molecular weights $1.6{\times}10^6\;dal\;and\;2.0{\times}10^6$ dal. The small plasmid (pSBK 112) specified macrolides, lincosamides and streptogramin type B (MLS) resistance gene which are expressed constitutively. Lage plasmid (pSBK 125) specified chloramphenicol resistance gene which is inducible. Strain 10-5 conferred a $3.0{\times}10^6$ dal plasmid (pSBK 141) which carry an inducible ampicillin resistance gene and strain P-H-2 conferred and $1.6{\times}10^6$ dal plasmid (pSBK 190) which carry a constitutive MLS resistance gene. Strain D-H-1 conferred four plasmids of molecular weights $0.8{\times}10^6$ dal (pSBK 201), $1.6{\times}10^6$ dal (pSBK 202), $2.5{\times}10^6$ dal (pSBK 203), and $1.2{\times}10^7$ dal (pDBK 204), respectively. Among those four plasmids, only pSBK 203 specified chloramphenicol resistance gene. Curing of constitutive MLS resistance using acriding orange or ethidium bromide in 2-23-6 and P-H-2 strains produced 'inducible' MLS resistance strains which are less resistant to MLS than the wild type strains, suggesting that there are two resistance genes in both strains; one is constitutive and the other is inducible.

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Studies on antibiotics resistance gene in Staphylococcus aureun Plasmid: Cloning of chloramphenicol resistance determinant (Staphylococcus aureus에서 분리된 plasmid상의 항생물질 저항성 인자에 관한 연구 : Chloramphenicol 저항성 인자의 클로닝)

  • 권동현;김영선;변우현
    • Korean Journal of Microbiology
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    • v.24 no.4
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    • pp.341-351
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    • 1986
  • R-plasmid(pSBK203, 2.5Mdal) conferring chloramphenicol resistance was isolated from mutiple antibiotic resistant Staphylococcus aureus D-H-1. Bacillus subtilis BD170 was transformed by this plasmid and restriction enzyme clevage sites of this plasmid were mapped for the cloning of chloramphenicol resistance gene. Taq I partial digested fragment of pSBK203(1.3kb) inserted into Cla I site of pBD9 appears to have both regulatory region for induction and structural gene for chloramphenicol resistance whereas Rsa I fragment (1.3kb, both ends are staggered away 0.1Kb from those of Taq I fragment) inserted into Sca I site of pBR322 showed constitutive expression in E. coli. Hinf I, Taq I, and Bgl II restriction enzyme recognition sites are found in both Rsa I fragment and Taq I fragment. Among these, Bgl II recognition site was associated with chloramphenicol resistance.

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