The Rep20 Replication Initiator From the pAG20 Plasmid of Acetobacter aceti

Abstract

In the previously isolated pAG20 plasmid from the Acetobacter aceti CCM3610 strain, the Rep20 protein was characterized as a main replication initiator. The pAG20 plasmid origin was localized in the vicinity of the rep20 gene and contained two 21-nucleotide-long iteron sequences, two 13-nucleotide-long direct repeats, and a DnaA-binding site. Electrophoretic mobility shift assay and nonradioactive fragment analysis confirmed that the Rep20 protein interacted with two direct repeats (5'-TCCAAATTTGGAT'-3') and their requirement during plasmid replication was verified by mutagenesis. Although the association could not be validated of the DnaA protein of from the host cells of Escherichia coli with the plasmid-encoded replication initiator that usually occurs during replication initiation, Rep20 was able to form dimeric structures by which it could bind the sequence of the rep20 gene and autoregulate its own expression. Targeted mutagenesis of the Rep20 protein revealed the importance of the third α-helix and ⁶³Lys, specifically during DNA binding. The second, closely adjacent β-sheet also took part in this process in which ⁵²Asn played a significant role.

Characterization of 12 Novel Microsatellite Markers ofSogatella furcifera(Hemiptera: Delphacidae) Identified From Next-Generation Sequence Data

“…2010 ). Although ISSR methods offer a relatively easy and inexpensive access to molecular genetic study ( Cichorz et al . 2014 ), yet several researchers doubts about their reproducibility, ascendancy, and homology ( Hansen et al .…”

Genomics of microbial plasmids: classification and identification based on replication and transfer systems and host taxonomy

“…Nevertheless, a well-defined method for the classification of Acetobacter plasmids has not been developed. Two groups have recently reported the characterization of Rep proteins, Rep7 and Rep20, from small plasmids pGR7 (Grones and Grones, 2012 ) and pAG20 (Babic et al, 2014 ), respectively, found in Acetobacter (class Rhodospirillales ). The Rep20 gene was also found in plasmids in Rhodospirillales and Rhodobacteriales (Figure 3C ).…”

Constitutive expression of a novel antimicrobial protein, Hcm1, confers resistance to both Verticillium and Fusarium wilts in cotton

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“…These results showed that the spread of V. dahliae was effectively hindered. A synthetic chimera of cecropin A and melittin CAPs with antimicrobial properties, MsrA1, effectively restricts Alternaria brassicae and Sclerotinia sclerotiorum infection in transgenic Brassica juncea and Solanum tuberosum plants 34 56 . The novel cecropin A-melittin hybrid peptide, CEMA, which has strong antimicrobial activity in vitro , confers resistance against Fusarium solani in transgenic tobacco 57 .…”

“…This chimeric protein showed a better antimicrobial spectrum than cecropin A alone, and less hemolytic activity than melittin alone 27 30 33 . The hybrid peptide can effectively inhibit the proliferation of pathogens in plants 33 34 35 , however, few hybrid peptides have been shown to activate plant innate immunity.…”