• 제목/요약/키워드: Cryptic plasmid

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Escherichia coli와 Corynebacterium glutamicum간의 shuttle vectors의 C. glutamicum에서의 안정성에 대한 클론된 유전자의 영향 (Effects of Cloned Genes on the Stability of Shuttle Vectors between Escherichia coli and Corynebacterium glutamicum)

  • 노갑수;김성준;오종원;이현환;현형환;이재흥
    • 미생물학회지
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    • 제29권3호
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    • pp.149-154
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    • 1991
  • Escherichia coli/Corynebacterium glutamicum shuttle vectors, pECCG1 and pECCG2 were constructed by joining a 3.00 kb cryptic plasmid pCB 1 from C. glutamicum and a 3.94 kb plasmid pACYC 177 from E. coli. By trimming unessential parts and introducing mulitiple cloning site into the plasmid pECCG 1, a plasmid pECCG122(5.1kb) was constructed. All the shuttle vectors were stably maintained in C. glutamicum up to about 40 generations irrespective of kanamycin addition in the medium. Threonine operon (homoserine dehydrogenase/homoserine kinase) and dapA gene (dihydrodipicolinate synthetase) of C. glutamicum were cloned into the plasmid pECCG122, and the resultant plasmids were designated pTN31 and pDHDP19, respectively. They were used to study the effect of cloned foreign gene on the stability of the plasmid pECCG122. Plasmids pTN31 and pDHDP19 were segregated rapidly from C. glutamicum when cultured in the medium without kanamycin. In medium with $50\mu${\g/ml} of kanamycin, their segregation rates were much slower than those in medium without kanamycin, but the danamycin addition didn't guarantee the complete maintenance of the plasmids in C. glutamicum.

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Characterization of Two Cryptic Plasmids from Levilactobacillus zymae GU240

  • Le, Huong Giang;Kim, Min Jae;Jeon, Hye Sung;Yoo, Ji Yeon;Kang, Yun Ji;Kim, Tae Jin;Kim, Jeong Hwan
    • 한국미생물·생명공학회지
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    • 제50권1호
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    • pp.63-70
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    • 2022
  • Two small cryptic plasmids, pHG1 and pHG2, were isolated from Levilactobacillus zymae (formerly Lactobacillus zymae) GU240 and characterized. pHG1 is 1,814 bp in size with a GC content of 37.4% and contains two open reading frames. orf1 can potentially encode a protein of 101 amino acids (aa) with 99% identity with the copy number control protein of Lacticaseibacillus paracasei. orf2 can potentially encode a protein of 230 aa with 99% identity with a replication protein from multiple species. Six inverted repeats (IR I-VI) and six direct repeats (DR I-VI) were found in pHG1. pHG2 is 2,864 bp in size, with a GC content of 39.6%. pHG2 has two orfs. orf1 might encode a protein with 99% identity with the TrsL transmembrane protein. orf2 might encode a protein with 99% identity with plasmid recombination proteins from lactic acid bacteria. Both pHG1 and pHG2 may be useful as frames for constructing lactic acid bacteria-Escherichia coli shuttle vectors.

pVC, a Small Cryptic Plasmid from the Environmental Isolate of Vibrio cholerae MP-1

  • Zhang, Ruifu;Wang, Yanling;Leung, Pak Chow;Gu, Ji-Dong
    • 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.

Pseudomonas nitroreducens TX1에 존재하는 작은 플라스미드의 특성 규명 (Characterization of a Small Cryptic Plasmid from Pseudomonas nitroreducens Strain TX1)

  • ;이경;강주범;황설이
    • 미생물학회지
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    • 제50권3호
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    • pp.210-215
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    • 2014
  • Pseudomonas nitroreducens TX1는 대만의 벼를 재배하는 논의 배수구에서 분리된 세균이다. 이 균주는 알킬페놀 폴리에톡실레이트와 같은 비이온성 계면활성제를 고농도에서도 탄소원으로 이용할 수 있다. 본 연구에서는 TX1 균주에서 분리된 새로운 플라스미드 pTX1의 특성을 조사하였다. 크기는 2,286 bp, GC 함량은 63.3%, 암호된 유전자로는 $Rep_{pTX1}$과 기능이 밝혀지지 않은 ORF1과 ORF2가 동정되었다. $Rep_{pTX1}$은 롤링-서클 기작에 의해 복제되는 그람 양성 세균에서 주로 발견되는 pC194/pUB110 플라스미드 계열에 속하는 DNA 복제 효소임을 알 수 있었다. 또한 세포마다 약 150개의 플라스미드가 존재함을 규명하였다. 플라스미드에 존재하는 유전자 지문과 유사 플라스미드와의 핵산과 아미노산 서열비교를 통해 pTX1은 슈도모나스 세균에서는 흔히 발견되지 않는 롤링-서클 기작에 의해 복제된다는 것을 확인할 수 있었다.

Screening of a Novel Lactobacilli Replicon from Plasmids of Lactobacillus reuteri KCTC 3678

  • Moon, Gi-Seong;Lee, Young-Duck;Kim, Wang-June
    • Food Science and Biotechnology
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    • 제17권2호
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    • pp.438-441
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    • 2008
  • A novel lactobacilli replicon from plasmids of Lactobacillus reuteri KCTC 3678 was isolated. Eight L. reuteri strains from Korean Collection for Type Cultures (KCTC) and Korea Food Research Institute (KFRI) were screened for cryptic plasmids and most strains harbored 1 or 2 plasm ids. Particularly, L. reuteri KCTC 3678 contained 6 plasm ids which all were used for screening of lactobacilli replicon. EcoRI digests of the plasmid DNA prep from L. reuteri KCTC 3678 were ligated with pUC19 and the recombinant DNAs were serially named from pLR1 to pLR7. A cat (chloramphenicol acetyltransferase; $Cm^r$) gene originated from pC194 was introduced into pLR1-7, resulting in pLR1cat-pLR7cat, respectively. The recombinant plasmids were introduced into L. reuteri KCTC 3679, and only transformants harboring pLR5cat were obtained, indicating that the insert in pLR5 functioned as a lactobacilli replicon.

유류분해균 Providencia rettgeri 4A3의 분리 및 원유분해 특성 (Isolation of Oil-Degrading Bacterium, Providencia rettagei 4A3 and Characterization of Crude Oil Degradation)

  • 김상우;유주순;이상철;조영수;이영춘
    • 생명과학회지
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    • 제11권6호
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    • pp.530-536
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    • 2001
  • 원유분해능이 강력한 해양균주를 얻고자 유류오염 지역 으로부터 crude oil을 탄소원으로 이용하는 십수종을 분리하였다. 분리된 균주중 원유분해 및 성장속도면에서 우수한 균주를 선별하여 형태학적, 생화학적 및 생리학적 특성을 조사한후 Providencia rettgeri 4A3로 동정하였다. 4A3균주의 원유분해를 위한 최적 배양조건은 배양온도 26$^{\circ}C$로 비교적 낮은 온도였으며, 초기 pH는 7.0이었다. 또한, 이 균주는 2.0% 염분농도에서 최대의 성장을 보여주어 해양 유래의균주임을 확인하였다. 4A3균주의 유화활성은 26$^{\circ}C$, pH 7.0의 배양조건으로 배양 3일째에 가장 높게 나타났으며, 유류분해의 경시적인 변화패턴을 관찰하였다. 분리균주는 약 7.0kb의 크기를 나타낸 미지의 플라스미드 1개를 보유하고 있었다.

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돼지유래 대장균의 항균제내성 분포와 R-plasmid의 성상 (Distribution of antimicrobial resistances and properties of R-plasmids in E coli isolated from pigs)

  • 정명은;여상건
    • 대한수의학회지
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    • 제34권4호
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    • pp.759-768
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    • 1994
  • E coli strains isolated from pigs were investigated with respect to antimicrobial resistances and prevalence of R-plasmids. Also determined were properties of R-plasmids by plasmid conjugation, curing and southern hybridization using gene probes. All of 400 E coli strains were resistant to CL and SU, and 0.3% to 96.8% of the strains were resistant to most antimicrobials such as TC, PG, AM, SM, CP, GM, EM, NM, etc, while all strains were sensitive to AK. All strains were also multiply resistant to three to twelve antimicrobials. The resistances to PG, SM, TC, AM, CP, SU and ST were transferable and supposed to be mediated by R-plasmids which were opportunistic for transposition into chromosome. Plasmids bigger in size than chromosomal DNA were considered as R-plasmids and most plasmids in small size (<4Kb) proved as cryptic plasmids or nonconjugative R-plasmids. In a strain(No 99), AM resistant property was determined from both chromosomal DNA and R-plasmid DNA which is bigger in size than chromosome.

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