Characterization of pLP18, a novel cryptic plasmid of Lactobacillus plantarum PC518 isolated from Chinese pickle

Pan, Qu; Zhang, Lin; Li, JinChuan; Chen, Tian; Chen, Wei; Wang, GuangKe; Yin, Jianhua

doi:10.1016/j.plasmid.2011.01.001

Cited by 9 publications

(7 citation statements)

References 34 publications

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“…To date, 22 species of Lactobacillus have been identified that contain plasmids. Lactobacillus plasmids vary widely in size (from 1.81–242.96 kb), number (from 1–10 different plasmids in a single strain), and gene content [ 6 , 14 , 15 ].…”

Section: Diversity and Similarity Of Plasmids From Labmentioning

confidence: 99%

“…Many small plasmids from L. plantarum replicate via the rolling-circle replication (RCR) mechanism, while some large plasmids, for example pMD5057, pWCFS103, and pST-III, are predicted to replicate via the theta mechanism [ 15 , 22 , 25 , 28 , 29 , 30 , 31 , 32 , 33 ]. Studies indicated that a rolling-circle plasmid pMRI 5.2 from a potentially probiotic strain L. plantarum BFE 5092, had two different plasmid-encoded replication initiation proteins from different replicon families, i.e.…”

Section: Diversity and Similarity Of Plasmids From Labmentioning

confidence: 99%

See 1 more Smart Citation

Plasmids from Food Lactic Acid Bacteria: Diversity, Similarity, and New Developments

Cui

et al. 2015

IJMS

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Plasmids are widely distributed in different sources of lactic acid bacteria (LAB) as self-replicating extrachromosomal genetic materials, and have received considerable attention due to their close relationship with many important functions as well as some industrially relevant characteristics of the LAB species. They are interesting with regard to the development of food-grade cloning vectors. This review summarizes new developments in the area of lactic acid bacteria plasmids and aims to provide up to date information that can be used in related future research.

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Section: Diversity and Similarity Of Plasmids From Labmentioning

confidence: 99%

Section: Diversity and Similarity Of Plasmids From Labmentioning

confidence: 99%

Plasmids from Food Lactic Acid Bacteria: Diversity, Similarity, and New Developments

Cui

et al. 2015

IJMS

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“…1), indicating that L. plantarum PC518 contains multiple plasmids. Among which, plasmid pLP18 with a size of 1.8 kb had been sequenced and published previously by ourselves [16]. Compared with the past report, the number of bands on gel increased from four to seven because the method of plasmid extraction had been improved.…”

Section: Resultsmentioning

confidence: 99%

“…A 150-bp fragment of pLP60 was amplified using primers pq15-F (5′- CCCCATAACACCCTACCT-3′) and pq15-R (5′- AAGACATTCTATCCCGAGT-3′), and a 170-bp fragment of pLP60 was amplified using primers pq17-F (5′-CTAAAGCGTCAACCGTAT-3′) and pq17-R (5′-TTCGTTCCCTTCGAGATG-3′). The gene of L-lactate dehydrogenase ( ldhL ) was used as the reference gene, and a 152-bp fragment of the ldhL gene was amplified [16]. The copy number of pLP60 was calculated using N relatives = (1 + E) − Δ C T [18].…”

Section: Methodsmentioning

confidence: 99%

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Detection and characterization of a theta-replicating plasmid pLP60 from Lactobacillus plantarum PC518 by inverse PCR

Yao

et al. 2019

Heliyon

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Plasmid DNA of Lactobacillus plantarum PC518 was isolated by an improved method which contained a washing step for removing lysozyme. Three plasmid DNA libraries were constructed. A pair of outward primers was designed at both ends of the novel plasmid fragment obtained from plasmid DNA libraries, and the remainder of the circle plasmid was amplified by inverse PCR (iPCR). The whole sequence of plasmid was analyzed by the basic local alignment search tool, Tandem Repeats Finder, DNAMAN V6.0, DNASTAR and MEGA X software. The copy number was measured using quantitative real-time PCR. Plasmid extract showed 7 bands on agarose gel, indicating that L. plantarum PC518 contains multiple plasmids. The complete sequence of plasmid pLP60 was obtained by plasmid DNA libraries and iPCR. pLP60 is 6006 bp in length with a G + C content of 41.19 %, which encodes 8 open reading frames (ORFs). The ori site like theta-type could be located upstream of repB , which contains a short tandem repeats (sTR) and a long tandem repeats (lTR). RepB of pLP60 only had low similarity with Rep protein of known theta-type plasmids, but phylogenetic tree analysis showed that plasmids whose Rep proteins are similar to pLP60 have lTR at ori , and the conservativeness of lTR is consistent with similarity of Rep proteins, suggesting that RepB of pLP60 is a theta-replicating protein. So pLP60 was classified as class A of theta replication. The copy number of pLP60 was measured as 5 copies per cell by qPCR.

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The dyeing effect of acridine orange for multiple plasmid systems is sensitive to temperature

Jiao,

Zhang,

Xie

et al. 2023

J of Cellular Biochemistry

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The Goldview dyeing of the natural multiplasmid system of Lactobacillus plantarum PC518 was affected by temperature. The article want to identify the specific molecules that cause temperature sensitivity, then experiment on the universality of temperature sensitivity, and finally preliminarily analyze the influencing factors. At 5°C and 25°C, single pDNA, multiplasmid system, and linear DNA samples were electrophoretic on agarose gel prestained by Goldview 1, 2, 3, and acridine orange (AO), respectively. Eighteen vectors of Escherichia coli and two vectors shortened by cloning were mixed into multiplasmid systems with different member numbers, and then electrophoresis with AO staining was performed within the range of 5°C–45°C, with a linearized multiplasmid system as the control. The lane profiles (peaks) were captured with Image Lab 5.1 software. After electrophoresis, the nine‐plasmid‐2 system was dyed with AO solutions of different ionic strengths to detect the effect of ionic strength on temperature sensitivity. It was measured that the UV‐visible absorption spectra of the nine‐plasmid‐2 system dissolved in AO solutions with different ionic strengths and pH. Further, a response surface model was constructed using Design‐Expert.V8.0.6 software. The electrophoresis result showed that the multiplasmid system from L. plantarum PC518 stained by AO staining showed a weak band at 5°C and five bands at 25°C, which was similar to the result of staining with Goldview 1, 2, and 3. The synthetic nine‐plasmid‐1 system and nine‐plasmid‐2 system displayed different band numbers on the electrophoresis gel in the electrophoresis temperature range of 5°C–45°C, namely 3, 4, 6, 4, and 2 bands, as well as 2, 6, 7, 8, and 5 bands. Using the 1× Tris‐acetate‐EDTA (TAE)‐AO solution, the poststaining results of the nine‐plasmid‐2 system in the temperature range of 5°C–45°C were 4, 6, 9, 9, and 7 bands, respectively. Further, using 5×, 10×, or 25× TAE buffer, the AO poststaining results at 5°C were 4, 2, and 1 bands, respectively. The ultraviolet spectral results from 5°C to 25°C showed that there was a significant difference (3.5 times) in the fluctuation amplitude at the absorption peak of 261.2 nm between 0× and 1–10× TAE‐AO solution containing the nine‐plasmid‐2 system. Specifically, the fluctuation amplitudes of 0×, 1×, 5×, and 10× samples were 0.032, 0.109, 0.112, and 0.110, respectively. At the same time, using 1× and 10× TAE buffer, the AO‐stained linear nine‐plasmid‐2 system remained stable and did not display temperature sensitivity. The response surface models of the AO‐stained nine‐plasmid‐2 system intuitively displayed that the absorbance of the 1× TAE samples increased significantly with increasing temperature compared to the 0× TAE samples, regardless of the pH value. The findings confirmed a temperature‐dependent effect in AO staining of natural or synthetic multiplasmid systems, with the optimum staining result occurring at 25°C. Ion strength was a necessary condition for the temperature sensitivity mechanism. This study layed the groundwork for further investigation into the reasons or underlying mechanisms of temperature sensitivity in AO staining of multiplasmid systems.

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Characterization of pLP18, a novel cryptic plasmid of Lactobacillus plantarum PC518 isolated from Chinese pickle

Cited by 9 publications

References 34 publications

Plasmids from Food Lactic Acid Bacteria: Diversity, Similarity, and New Developments

Plasmids from Food Lactic Acid Bacteria: Diversity, Similarity, and New Developments

Detection and characterization of a theta-replicating plasmid pLP60 from Lactobacillus plantarum PC518 by inverse PCR

The dyeing effect of acridine orange for multiple plasmid systems is sensitive to temperature

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