Seamless insert-plasmid assembly at sub-terminal homologous sequences
Publication date: Available online 24 October 2019Source: PlasmidAuthor(s): Anna-Sophia Krebs, Tobias Bierig, Gabriella Collu, Roger M. BenoitAbstractThe engineering of fusion proteins for structural biology and protein nanotechnology often requires seamless DNA assembly with slight variations in the domain boundaries. To improve the molecular biology workflow for such projects, we evaluated the use of sub-terminal homologous sequences (HS) for co-transformation cloning and for T5 exonuclease / Phusion DNA polymerase mediated in vitro assembly. To quantify the effects of different HS-to-ends distances on cloning efficiency...
Source: Plasmid - October 26, 2019 Category: Biotechnology Source Type: research
The biology of IncI2 plasmids shown by whole-plasmid multi-locus sequence typing
Publication date: Available online 17 October 2019Source: PlasmidAuthor(s): Richard J. MeinersmannAbstractIncI2 type plasmids are medium-sized (~55–80 kb) conjugative plasmids that have been found carrying important antimicrobial resistance genes but have also been frequently found as cryptic plasmids. The DNA sequences for 147 fully sequenced IncI2 plasmids were studied by a whole-plasmid multi-locus sequence typing (wpMLST) scheme. A total of 171 loci were identified of which 52 were considered core (carried by greater than 95% of the plasmids). Most of the plasmids carrying the antimicrobial gene mcr-1 were in a dis...
Source: Plasmid - October 19, 2019 Category: Biotechnology Source Type: research
Identification of a novel conjugative plasmid carrying the multiresistance gene cfr in Proteus vulgaris isolated from swine origin in China
In this study, a novel conjugative plasmid carrying cfr, designated as pPvSC3, was characterized in a Proteus vulgaris strain isolated from swine in China. Plasmid pPvSC3 is 284,528 bp in size and harbors 10 other antimicrobial resistance genes, making it a novel plasmid that differs from all known plasmids due to its unique backbone and repA gene. BLAST analysis of the plasmid sequence shows no significant homology to any known plasmid backbone, but shows high level homology to Providencia rettgeri strain CCBH11880 Contig_9, a strain isolated from surgical wound in Brazil, 2014. There are two resistance-determining regi...
Source: Plasmid - September 7, 2019 Category: Biotechnology Source Type: research
Editorial
Publication date: Available online 28 August 2019Source: PlasmidAuthor(s): Eva M. Top, Beth Traxler, Julian I. Rood (Source: Plasmid)
Source: Plasmid - August 29, 2019 Category: Biotechnology Source Type: research
Inside or outside? A new collection of Gateway vectors allowing plant protein subcellular localization or over-expression
Publication date: Available online 23 August 2019Source: PlasmidAuthor(s): François Berthold, David Roujol, Caroline Hemmer, Elisabeth Jamet, Christophe Ritzenthaler, Laurent Hoffmann, Corinne Schmitt-KeichingerAbstractTransient expression of proteins based on agro-infiltration techniques has proven very efficient and straightforward to study the intrinsic properties of proteins. The level of protein expression has been enhanced by the use of vector plasmids containing virus-derived sequences and the cloning step has been facilitated by recombination technologies. The pEAQ-HT-DEST series of vectors fulfilling these improv...
Source: Plasmid - August 25, 2019 Category: Biotechnology Source Type: research
Transformation of the rhizospheric Bacillus cereus sensu lato B25 strain using a room-temperature electrocompetent cells preparation protocol
Publication date: Available online 30 July 2019Source: PlasmidAuthor(s): EstefanÃa Morales-Ruiz, Astrid López Ceballos, Ignacio E. Maldonado-MendozaAbstractBacterial transformation is a crucial step in the genetic manipulation of a bacterium. However, Gram-positive bacteria are difficult to transform and consequently many different methodologies have been developed. Here, we examined the transformation efficiencies of an electroporation protocol by varying three main factors: the composition of the electroporation buffer, the strength of the electric pulse, and the composition of the recovery media. Overall, transformati...
Source: Plasmid - July 31, 2019 Category: Biotechnology Source Type: research
Editorial Board
Publication date: July 2019Source: Plasmid, Volume 104Author(s): (Source: Plasmid)
Source: Plasmid - July 26, 2019 Category: Biotechnology Source Type: research
Publisher's Note
Publication date: July 2019Source: Plasmid, Volume 104Author(s): (Source: Plasmid)
Source: Plasmid - July 26, 2019 Category: Biotechnology Source Type: research
B/O plasmid R16 from 1956 carries an In1-like class 1 integron embedded in a complex region containing parts of the Acinetobacter baumannii AbaR resistance island
Publication date: Available online 18 July 2019Source: PlasmidAuthor(s): Robert A. Moran, Ruth M. HallAbstractThe IncB/O multiresistance plasmid R16, recovered in 1956 and used in experimental studies of B/O plasmid features, was sequenced. The resistance genes are all within a class 1 integron closely related to In1 containing the cassette array oxa2-aadA1-oxa2-orfD and sul1 in the 3′-conserved segment (3′-CS), with Tn10, containing tetA(B), inserted just inside the 3′-CS. The integron is part of a 25,144 bp region inserted in the plasmid backbone, bounded on only one side by a truncated Tn6018 and flanked by a 4â...
Source: Plasmid - July 18, 2019 Category: Biotechnology Source Type: research
Improving biosynthetic production of pinene through plasmid recombination elimination and pathway optimization
Publication date: Available online 15 July 2019Source: PlasmidAuthor(s): Shao-Heng Bao, Dong-Yi Zhang, Er MengAbstractPinene is a monoterpene with wide industrial applications, especially as a promising high energy-density jet fuel. Traditional production of pinene on an industrial scale is material consumptive and has a low yield. As an alternative, microbial organisms have been engineered though advanced synthetic biological techniques to produce a variety of heterologous products, including pinene. Here, we investigated the stability of genetic circuits encoding the pinene producing pathway during fermentation and its r...
Source: Plasmid - July 16, 2019 Category: Biotechnology Source Type: research
The piggyBac-based double-inducible binary vector system: A novel universal platform for studying gene functions and interactions
Publication date: Available online 29 June 2019Source: PlasmidAuthor(s): Wencui Sun, Jiawen Teng, Jiahui Zeng, Yuan Xue, Jing Chang, Yonggang Zhang, Xu Pan, Ya Zhou, Mowen Lai, Guohui Bian, Qiongxiu Zhou, Jiaxin Liu, Bo Chen, Feng MaAbstractEukaryotic inducible overexpression systems, including Tet-On and mifepristone-inducible systems, have been widely used to study gene functions by reverse genetics. Among the transposon systems reported to date, the piggyBac transposon system is one of the most efficient in cultured mammalian cells. Here, we report a piggyBac-based double-inducible system that combined the advantages of...
Source: Plasmid - June 30, 2019 Category: Biotechnology Source Type: research
Assembly of large mobilizable genetic cargo by double recombinase operated insertion of DNA (DROID)
Publication date: Available online 25 June 2019Source: PlasmidAuthor(s): Kevin Neil, Nancy Allard, David Jordan, Sébastien RodrigueAbstractThere is an important need to develop new therapeutic tools to modulate the gene content of microbiomes. A potential strategy for microbiome engineering relies on the delivery of genetic payloads by conjugative plasmids. Yet, the introduction of large DNA molecules in conjugative plasmids can be challenging. Here, we describe the Double Recombinase Operated Insertion of DNA (DROID), an efficient method to assemble large DNA molecules without introducing antibiotic resistance genes or o...
Source: Plasmid - June 25, 2019 Category: Biotechnology Source Type: research
A phylogenetic test of the role of CRISPR-Cas in limiting plasmid acquisition and prophage integration in bacteria
Publication date: Available online 11 June 2019Source: PlasmidAuthor(s): Derek O'Meara, Leonard NunneyAbstractCRISPR-Cas is a prokaryotic defense system capable of protecting the cell from damaging foreign genetic elements. However, some genetic elements can be beneficial, which suggests the hypothesis that bacteria with CRISPR-Cas incur a cost of reduced intake of mutualistic plasmids and prophage. Here we present the first robust test of this hypothesis that controls for phylogenic and ecological biases in the distribution of CRISPR-Cas. We filtered the available genomic data (~7000 strains from ~2100 species) by first s...
Source: Plasmid - June 12, 2019 Category: Biotechnology Source Type: research
Plasmid analysis of Escherichia coli isolates from South Korea co-producing NDM-5 and OXA-181 carbapenemases
Publication date: Available online 28 May 2019Source: PlasmidAuthor(s): Jin Yang Baek, Sun Young Cho, So Hyun Kim, Cheol-In Kang, Kyong Ran Peck, Jae-Hoon Song, Doo Ryeon Chung, Kwan Soo KoAbstractRecently, Escherichia coli isolates co-producing New Delhi metallo-β-lactamase (NDM)-5 and oxacillinase (OXA)-181 were identified in a tertiary-care hospital of South Korea. Isolate CC1702–1 was collected from urine in January 2017 and isolate CC1706-1 was recovered from a transtracheal aspirate of a hospitalized patient in May 2017. Carbapenemase genes were identified by multiplex PCR and sequencing, and whole genome sequenci...
Source: Plasmid - May 30, 2019 Category: Biotechnology Source Type: research
Editorial Board
Publication date: May 2019Source: Plasmid, Volume 103Author(s): (Source: Plasmid)
Source: Plasmid - May 18, 2019 Category: Biotechnology Source Type: research
