• 미생물학회지
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• 제27권4호
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• pp.334-341
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• 1989

Toluate 분해 플라스미드를 pseudomonase cepacia SUB37에서 분리하여 분자량은 한천 젤 전기영동으로 측정한 결과 79. (119kb)로 확인되었다. 이 TOL플라스미드는 Pseudomonas의 다른 균주와 다른 속의 균주에 전달되었다. m-toluate 분해에서 가장 중요한 역할을 하는 catechol-2,3-oxygenase 활성을 P. cepacia SUB37과 transconjugant의 조효소액으로부터 측정한 결과, P. putida mt 2에서와 같이, meta pathway를 거쳐 m-toluate를 분해하는 유전자들이 plasmid에 암호화됨을 알수 있었다. P. cepacia SUB37 유래의 새로운 TOL plasmid는 IncP-4 불화합성군에 속하였고, 이것은 아마도 P. putida의 IncP-9 그룹의 TOL 플라스미드의 유도체로 사료된다.

• 한국미생물·생명공학회지
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• 제18권4호
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• pp.433-436
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• 1990

접합에 의한 P.putida mt-2의 TOL를 CAM 함유 P.putida PpG1으로 이동시켜 형성된 접합주 P.Putida CST3A는 작아진 TOL(TOL$\Delta$)를 가지고 있었지만, m-toiuate를 분해할 수 있었다. 접합에 의한 이동실험은 CAM에 결합되어 CAM:TO* 플라스미드를 형성하고 있었다. 불화합성 Inc P9군에 속하는 NAH를 CAM:TOL* 과 TOL$\Delta$을 가지는 P.putida CST3A로 이동시키면 TOL$\Delta$의 방출이 관찰되었으나 m-toluate 대사에는 아무런 영향을 미치지 않았다.

• 미생물학회지
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• 제25권2호
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• pp.87-93
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• 1987

The properties of the plasmid pKU41 isolated from Pseudomonas putkda KU190 have been investigated, pKU41 was defined as an R plasmid having a transmissible ampicillin and tetracycline resistance determinant, and could be classified as a plasmid belonging to IncP-1 group according to incompatibility grouping.

• 대한수의학회지
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• 제29권2호
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• pp.59-67
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• 1989

This paper deals with the genetic properties of R plasmids in Salmonella originated from pigs and cattle. The plasmid DNA was examined for incompatibility, stability and fertility inhibition(Fi), and gel electrophoresis was performed for isolation of plasmid DNA. The results obtained were summerized as follows: 1. Among the 66 conjugative R plasmids from 44 pigs and 22 cattle, 61 R plasmids (92.4%) were $Fi^-$, whereas the remainder were $Fi^+$. 2. The Inc groups of 66 R plasmids were determined with 7 standard plasmids. Twenty-six R plasmids were classified into Inc group $I{\alpha}$, H1, H2 or F1, 40 R plasmids being not classified with standard plasmids used, and the Inc group $I{\alpha}$ (57.7%) was most frequent. 3. Inc groups $I{\alpha}$, H1, and F1 were identified in strains from swine, Inc groups H2 and F1 from cattle. 4. The plasmid DNA profiles in 16 Salmonella isolated from pigs and cattle were confirmed as being 1 to 10 fragments by the gel eletrophoresis. Their molecular weight ranged 1.0 to 90 megadalton. 5. The molecular weight of conjugative plasmids ranged 1.0 to 80 megadalton in 4 Salmonella (P-4, P-5, P-7 and P-8) isolated from pigs.

• Journal of Microbiology and Biotechnology
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• 제17권1호
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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.

• 미생물학회지
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• 제31권1호
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• pp.37-43
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• 1993

The effect of stb locus of E. COLI IncFII plasmid NR1 on the stability of chimeric plasmids was investigated. First, we have isolated the stability locus (stb) from E. coli NR1 plasmid and then inserted into the three different vectors, pUC8, YRp17 and YEp24. By examining their stability in E. coli and yeast, we showed that the recombinant plasmids containing stb locus were resonably stable. Also, by comparing the amounts of the rDNA fragments per haploid genome with those of the plasmid fragments, we showed they copy number of recombinant plasmids was not increased. Consequently, the stb locus of E. coli IncFII plasmid NR1 stabilized the chimeric plasmids but did not affect the replication or copy number of plasmids.

• Journal of Microbiology and Biotechnology
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• 제21권11호
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• pp.1123-1126
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• 2011

The IncP plasmid R995 has been a useful self-transmissible, broad-host-range vector for a number of applications including the recombinase/conjugation-based cloning of large genomic DNA segments. However, R995 derivatives (or related plasmids) expressing a wide range of different resistance markers and Flp recombinase target sites do not exist in the literature. In addition, documented strategies for applying such plasmids in cloning applications that take advantage of conjugation for the convenient isolation and recovery of constructs are extremely limited. Here, we report a new series of R995 plasmids with alternative markers to increase options for applications in backgrounds already expressing resistance to a particular antibiotic(s). These R995 plasmids have been engineered to contain FRT sites that can be used for recombinase-based cloning. We demonstrate the utility of this approach by cloning 20 kb regions from the Salmonella Typhimurium and Escherichia coli genomes and by cloning DNA from an exogenous plasmid source. To our knowledge, this represents the first systematic engineering of an intact, self-transmissible IncP plasmid with a series of alternative antibiotic markers and FRT sites.

• 미생물학회지
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• 제25권4호
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• pp.282-289
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• 1987

Pseudomonas putida KU816에서 분리한 플라스미드 pKU10의 여러가지 특성을 조사하고 그 제한 효소 지도를 작성하였다. pKU 10은 암펴설런, 테트라사이클린, 클로람페니콜에 대한 내성 유전자를 갖는 작은 R factor로서 마이토마이신 C에 의하여 큐어링 된다. 플라스미드의 크기는 9.4Kb로 측정되었다. pKU 10은 Pseudomonas와 E.coli블 숙주로 하였을 때 안정하게 형질 발현이 되다. 또한 pKU 10의 불화합성균은 IncP-I으로 조사되었다. Eco RI, Xho I. SaiI, BglII, SmaI은 pKU 10 DNA를 한 부위에서 자르고, Pst I은 두 부위, Hind Ill는 여섯 부위에서 자른다. 제한 효소 지도는 제한 효소를 이중, 삼중으로 완전 소화시키거나, 부분 소화시켜서 얻었다. pKU 10은 Pseudomonas속에서 유용한 클로닝 벡터호 이용된 것으로 기대된다.

• Journal of Microbiology and Biotechnology
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• 제20권5호
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• pp.871-874
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• 2010

Many studies require expression analysis of the same gene/promoter across a range of bacterial genera. However, there is currently a lack of availability of reporters based on the broad-host-range IncQ replicon, which is compatible with a popular improved IncP transfer system that is self-transfer defective. We report IncQ lacZ reporter plasmids with features including (1) compatibility with IncP, IncW, and pBHR/pBBR replicons, (2) a variety of antibiotic markers (Sp-r, Sm-r, Km-r, Cm-r), (3) convenient mobilization via a novel self-transfer-defective IncP conjugation system, and (4) GenBank DNA sequences. Utility is demonstrated using three different promoters in different Gram-negative genera.

• 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.