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Isolation of a minireplicon of the plasmid pG6303 of Lactobacillus plantarum G63 and characterization of the plasmid-encoded Rep replication protein

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Abstract

A cryptic 10.0-kb plasmid pG6303 from a multiplasmid-containing Lactobacillus plantarum G63 was studied. The analysis of replicon was facilitated by the construction of shuttle vectors and electrotransformation into L. plantarum. The pG6303 replicon included (i) an open reading frame encoding the putative Rep replication initiation protein; and (ii) the putative origin of replication. The Rep protein was expressed as a fusion with the hexa-histidine (His) at its C-terminal end and purified by Ni-affinity chromatography. The electrophoretic mobility shift assays in pG6303 showed that the purified Rep protein specifically bound from 5582 to 5945 bp, differing from the putative origin of replication of pG6303. We speculate that pG6303 replication is a new mode of plasmid replication.

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Acknowledgement

This work was financially supported by the National High-Tech Development Project (2011AA100805-1).

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Authors and Affiliations

  1. Key Laboratory of Agricultural Environmental Microbiology, Ministry of Agriculture, College of Life Sciences, Nanjing Agricultural University, Nanjing, Jiangsu, 210095, People’s Republic of China

    JING FAN, XUEDONG XI, YAN HUANG & ZHONGLI CUI

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  1. JING FAN
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  2. XUEDONG XI
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  3. YAN HUANG
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  4. ZHONGLI CUI
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Correspondence to ZHONGLI CUI.

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[Fan J., Xi X., Huang Y. and Cui Z. 2015 Isolation of a minireplicon of the plasmid pG6303 of Lactobacillus plantarum G63 and characterization of the plasmid-encoded Rep replication protein. J. Genet. 94, xx–xx]

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FAN, J., XI, X., HUANG, Y. et al. Isolation of a minireplicon of the plasmid pG6303 of Lactobacillus plantarum G63 and characterization of the plasmid-encoded Rep replication protein. J Genet 94, 177–186 (2015). https://doi.org/10.1007/s12041-015-0500-6

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