• Genomics & Informatics
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• 제1권1호
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• pp.55-60
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• 2003

The nucleotide sequence of pZMO1, a small cryptic plasmid of Zymomonas mobilis ATCC10988 was determined. Analysis of 1,680 bp of sequence revealed $69\%$ identity with Shigella sonnei plasmid, pKYM and $61\%$ identity with Nostoc sp. ss DNA replicating plasmid. Analysis of a deduced amino acid sequence of an orf of pZMO1 revealed $75\%$ identity and $90\%$ similarity with the repA gene of Synechocystis sp. plasmid pCA2.4. The upstream region of the repA gene of pZMO1 possesses six directed repeat sequences and two inverted repeat sequences at downstream of the IR consensus sequence of nick region of rolling circle replication (RCR) plasmid. A typical terminator hairpin structure was found at the downstream region of repA gene. Degradation of single-stranded plasmid DNA by S1 nuclease was detected by Southern hybridization. It suggests that pZMO1 replicates by a rolling circle mechanism in Z. mobilis ATCC10988 cells.

• 산업기술연구
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• 제28권B호
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• pp.53-57
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• 2008

From the extensive screening for small cryptic plasmid among about 23 lactic acid bacteria (LAB), 2.4 kb of cryptic plasmid was isolated from Lactobacillus farciminis strain KCTC 3681 and named as pLF24. The plasmid pLF24 was a circular molecule of 2,396 base-pairs in length with a G+C content of 38%. Two protein-coding sequences could be predicted. ORF1 and ORF2 showed homologies to plasmids of gram-positive bacteria. The replication protein coded by ORF2 and the plus origin, were similar to replication regions of other gram-positive bacteria as shown in plasmids such as pLH2, pLS141-1 and pLC2. The nucleotide sequence of pLF24 was deposited into Genbank data base with an accession number of EU429343. The newly isolated plasmid can be used for construction of shuttle vector in Lactobacillus bacteria.

• BMB Reports
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• 제34권6호
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• pp.584-589
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• 2001

One of the cryptic plasmids from the oil degrading bacterium Klebsiella sp. KCL-2, the small plasmid pGD2, has been identified and characterized. This plasmid has a size of 3.6 kb with unknown functions. We constructed the recombinant plasmid pMGD2. The nucleotide sequences of the plasmid were determined and two open reading frames were detected. ORF1 encodes a replication initiator protein (RepA), which has a high degree of homology with the protein of ColE2 plasmid. The product encoded by ORF2 showed a high similarity with the transposase protein of IS5. IS5 is 1195 by long and contains an inverted terminal repetition of 16 bp with one mismatch. Stem-loop structures in the 5'untranslated region of the repA suggest that a putative gene, incA, is located in a complementary strand to the leader region of the repA mRNA.

• Journal of Microbiology
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• 제45권3호
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• pp.193-198
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• 2007

A marine bacterium was isolated from Mai Po Nature Reserve of Hong Kong and identified as Vibrio cholerae MP-1. It contains a small plasmid designated as pVC of 3.8 kb. Four open reading frames (ORFs) are identified on the plasmid, but none of them shows homology to any known protein. Database search indicated that a 440 bp fragment is 96% identical to a fragment found in a small plasmid of another V. cholerae. Further experiments demonstrated that a 2.3 kb EcoRI fragment containing the complete ORF1, partial ORF4 and their intergenic region could self-replicate. Additional analyses revealed that sequence upstream of ORF1 showed the features characteristic of theta type replicons. Protein encoded by ORF1 has two characteristic motifs existed in most replication initiator proteins (Rep): the leucine zipper (LZ) motif located at the N-terminal region and the alpha helix-turn-alpha helix motif (HTH) located at the C-terminal end. The results suggest that pVC replicates via the theta type mechanism and is likely a novel type of theta replicon.

• 약학회지
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• 제52권2호
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• pp.147-150
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• 2008

Previously, we reported on the antibiotic resistance patterns of 50 strains of Staphylococcus aureus which were isolated from a hospital in Busan, Korea from July 2005 to December 2006. We have isolated small plasmids and classified plasmid types by agarose gel electrophoresis. We have selected 5 plasmids and determined complete nucleotide sequences of those plasmids. The aim of this paper is to report on the characteristics of cadmium, erythromycin, lincomycin resistance plasmids and a cryptic plasmid based on the sequence analysis obtained by using the BLAST program.

• 약학회지
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• 제43권6호
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• pp.789-792
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• 1999

The complete nucleotide sequence of pKH4, a small multidrug resistance (smr) plasmid isolated from multidrug resistant Staphylococcus aureus SA5, was determined. Sequence analysis has revealed that pKH4 has two open reading frames for Rep and Smr proteins. The comparison of the amino acid sequence of Smr protein of pKH4 with those of other Smr proteins of various Staphylococcus showed that Smr protein of pKH4 is a new member of the SMR family.

• International Journal of Industrial Entomology and Biomaterials
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• 제9권2호
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• pp.183-186
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• 2004

Recently, we have developed an easy, simple and convenient circular DNA cloning system named plasmid capture system (PCS). To investigate usefulness of PCS in cloning of plasmids from Bacillus thuringiensis strains, PCS donors, pPCS-S and pPCS-L were applied to clone plasmids of B. thuringiensis subsp. israelensis by in vitro transposition using 4{TnsABC^*}$ transposase. In result, 3 small plasmids were cloned, and these were consistent with pTX14-1, pTX14-2 and pTX14-3 reported previously from B. thuringiensis subsp. israelensis. Therefore, the PCS can be successfully applied to clone small plasmids from B. thuringiensis strains.

• 미생물학회지
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• 제23권4호
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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.

• 한국미생물·생명공학회지
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• 제27권5호
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• pp.364-369
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• 1999

In order to obtain the suitable plasmids for constructing plasmid vectors of Bacillus species, endogeneous plasmid DNAs were screended from thermo-tolerant soil bacteria. Based on agarose gel electrophoresis patterns of the isolated plasmid DNAs, two strains harboring small-size plasmids were selected. The isolated were identified to belong to the genus Bacillus on the basis of their morphological and biochemical properties, and named Bacillus sp. 3-3 and 77-8, respectively. The restriction endonuclease maps were determined for four plasmids including two plasmids from each Bacillus isolates. It is interesting that Bacillus sp. 3-3 and 77-8 have an identical plasmid according to the restriction maps. The three kinds of hybrid plasmids constructed by introducing each plasmid of two isolates into a Escherichia coli plasmid vector. pUCCm18 containing chloramplenicol resistance gene active in Bacillus strains, could be replicated in B. subtilis and B. licheniformis. These plasmids are very stable in B. subtilis, suggesting that the Bacillus plasmids identified in this work would be useful for development of new cloning vectors for Bacillus strains.

• 미생물학회지
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• 제24권1호
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• pp.12-17
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• 1986

A new computer algorithm is described to order the restriction fragments of plasmid DNA which has been cleaved with several restriction endonucleases in single or double digestions rapidly with realistic error rates. The permutation and high weight on small fragments methods construct all logical circular map solutions. The program is written in Apple BASIC and run on an Apple II plus microcomputer with 64K memory. Several examples are presented which indicate the high efficiency of the profram in construction possible restriction map for YEp24.