IF 2.6 4区生物学

Plasmid Pub Date : 2021-07-01 DOI: 10.1016/j.plasmid.2021.102578

Milan S. Stosic , Marianne Sunde , Solveig Sølverød Mo , Amar Anandrao Telke , Knut Rudi

{"title":"Interference of ISEcp1-blaCTX-M-1 with the shufflon rearrangement in IncI1 plasmids","authors":"Milan S. Stosic , Marianne Sunde , Solveig Sølverød Mo , Amar Anandrao Telke , Knut Rudi","doi":"10.1016/j.plasmid.2021.102578","DOIUrl":"https://doi.org/10.1016/j.plasmid.2021.102578","url":null,"abstract":"<div>IncI1 plasmids are known disseminators of the extended-spectrum cephalosporin resistance (ESC) gene blaCTX-M-1, among species of the Enterobacteriaceae family. In several IncI1 plasmids, this gene was found incorporated into the transposition unit, ISEcp1-blaCTX-M-1-orf477, interrupting a shufflon region, a hallmark of IncI1 conjugative plasmids. The shufflon diversifies pilV gene that encodes the adhesine-type protein found on the tip of the conjugative pilus. To further elucidate the shufflon rearrangement, we examined to what extent the shufflon rearrangement was affected by the transposition-unit insertion. As expected, the interrupted shufflons generated a lower number of shufflon variants and exhibited an altered segment-deletion pattern compared to the non-interrupted shufflon. Interestingly, segment-loss frequency of the interrupted shufflons was distinctive in different plasmid hosts. Finally, the analysis of the 3′ end of the pilV gene revealed that shufflon rearrangement favoured segment A to complete pilV partial open reading frame regardless of the shufflon. Thereby, it could be assumed that the A-segment has greater importance during conjugation, however, this remained a hypothesis. Further exploration of shufflon rearrangement and its importance in the plasmid-recipient selection during conjugation would be beneficial as the knowledge could be applied in developing a strategy to limit IncI1 mediated antimicrobial resistance dissemination.</div>","PeriodicalId":49689,"journal":{"name":"Plasmid","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2021-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.plasmid.2021.102578","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"72286540","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 1

Utilization of a tetracycline-inducible system for high-level expression of recombinant proteins in Francisella tularensis LVS 利用四环素诱导系统在土拉弗朗西斯菌LVS中高水平表达重组蛋白

IF 2.6 4区生物学

Plasmid Pub Date : 2021-05-01 DOI: 10.1016/j.plasmid.2021.102564

Valeria Sheshko , Marek Link , Igor Golovliov , Lucie Balonova , Jiri Stulik

{"title":"Utilization of a tetracycline-inducible system for high-level expression of recombinant proteins in Francisella tularensis LVS","authors":"Valeria Sheshko , Marek Link , Igor Golovliov , Lucie Balonova , Jiri Stulik","doi":"10.1016/j.plasmid.2021.102564","DOIUrl":"10.1016/j.plasmid.2021.102564","url":null,"abstract":"<div>Francisella tularensis is a Gram-negative intracellular pathogen causing tularemia. A number of its potential virulence factors have been identified, but their biology and functions are not precisely known. Understanding the biological and immunological functions of these proteins requires adequate genetic tools for homologous and heterologous expression of cloned genes, maintaining both original structure and post-translational modifications. Here, we report the construction of a new multipurpose shuttle plasmid – pEVbr – which can be used for high-level expression in F. tularensis. The pEVbr plasmid has been constructed by modifying the TetR-regulated expression vector pEDL17 (LoVullo, 2012) that includes (i) a strong F. tularensis bfr promoter, and (ii) two tet operator sequences cloned into the promoter. The cloned green fluorescent protein (GFP), used as a reporter, demonstrated almost undetectable basal expression level under uninduced conditions and a highly dynamic dose-dependent response to the inducer. The utility of the system was further confirmed by cloning the gapA and FTT_1676 genes into the pEVbr vector and quantifying proteins expression in F. tularensis LVS, as well as by studying post-translational modification of the cloned genes. This study demonstrates that high levels of recombinant native-like Francisella proteins can be produced in Francisella cells. Hence, this system may be beneficial for the analysis of protein function and the development of new treatments and vaccines.</div>","PeriodicalId":49689,"journal":{"name":"Plasmid","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2021-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.plasmid.2021.102564","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25388357","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 1

The extrachromosomal elements of the Naegleria genus: How little we know Naegleria属的染色体外成分:我们所知甚少

IF 2.6 4区生物学

Plasmid Pub Date : 2021-05-01 DOI: 10.1016/j.plasmid.2021.102567

B.T. Nguyen , N.M. Chapman , S. Tracy , K.M. Drescher

引用次数: 2

An improved direct metamobilome approach increases the detection of larger-sized circular elements across kingdoms 一种改进的直接变微生物组方法增加了跨王国的较大尺寸圆形元素的检测

IF 2.6 4区生物学

Plasmid Pub Date : 2021-05-01 DOI: 10.1016/j.plasmid.2021.102576

Katrine Wacenius Skov Alanin , Tue Sparholt Jørgensen , Patrick Denis Browne , Bent Petersen , Leise Riber , Witold Kot , Lars Hestbjerg Hansen

{"title":"An improved direct metamobilome approach increases the detection of larger-sized circular elements across kingdoms","authors":"Katrine Wacenius Skov Alanin , Tue Sparholt Jørgensen , Patrick Denis Browne , Bent Petersen , Leise Riber , Witold Kot , Lars Hestbjerg Hansen","doi":"10.1016/j.plasmid.2021.102576","DOIUrl":"10.1016/j.plasmid.2021.102576","url":null,"abstract":"<div>Mobile genetic elements (MGEs) are instrumental in natural prokaryotic genome editing, permitting genome plasticity and allowing microbes to accumulate genetic diversity. MGEs serve as a vast communal gene pool and include DNA elements such as plasmids and bacteriophages (phages) among others. These mobile DNA elements represent a human health risk as they can introduce new traits, such as antibiotic resistance or virulence, to a bacterial strain. Sequencing libraries targeting environmental circular MGEs, referred to as metamobilomes, may broaden our current understanding of the mechanisms behind the mobility, prevalence and content of these elements. However, metamobilomics is affected by a severe bias towards small circular elements, introduced by multiple displacement amplification (MDA). MDA is typically used to overcome limiting DNA quantities after the removal of non-circular DNA during library preparations. By examining the relationship between sequencing coverage and the size of circular MGEs in paired metamobilome datasets with and without MDA, we show that larger circular elements are lost when using MDA. This study is the first to systematically demonstrate that MDA is detrimental to detecting larger-sized plasmids if small plasmids are present. It is also the first to show that MDA can be omitted when using enzyme-based DNA fragmentation and PCR in library preparation kits such as Nextera XT® from Illumina.</div>","PeriodicalId":49689,"journal":{"name":"Plasmid","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2021-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.plasmid.2021.102576","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38888999","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2

Reciprocal relation between reporter gene transcription and translation efficiency in fission yeast 裂变酵母报告基因转录与翻译效率的互反关系

IF 2.6 4区生物学

Plasmid Pub Date : 2021-05-01 DOI: 10.1016/j.plasmid.2021.102557

Suchita Srivastava , Satinderdeep Kaur , Hemant K. Verma , Suman Rani , Manisha Thakur , Swati Haldar , Jagmohan Singh

引用次数: 0

Molecular characterization of two novel NDM-1-producing atypical enteroaggregative Escherichia coli isolates from patients 两株新型产ndm -1的非典型肠聚集性大肠杆菌分离株的分子特征

IF 2.6 4区生物学

Plasmid Pub Date : 2021-05-01 DOI: 10.1016/j.plasmid.2021.102568

Pengcheng Du , Pei Zhang , Juan Wang , Ruichao Li , Séamus Fanning , Li Bai

{"title":"Molecular characterization of two novel NDM-1-producing atypical enteroaggregative Escherichia coli isolates from patients","authors":"Pengcheng Du , Pei Zhang , Juan Wang , Ruichao Li , Séamus Fanning , Li Bai","doi":"10.1016/j.plasmid.2021.102568","DOIUrl":"10.1016/j.plasmid.2021.102568","url":null,"abstract":"<div>To investigate NDM-1-producing atypical Enteroaggregative Escherichia coli (aEAEC) of sequence type 349 from hospitalized patients, the isolates 13ZX28 and 13ZX36 were subjected to antimicrobial susceptibility testing, conjugation and whole genome sequencing. Only one single nucleotide mutation was detected in chromosomes despite different plasmid profiles. Both isolates were positive for blaNDM-1 mediating resistance to carbapenem. A novel plasmid p13ZX28–272 (~272-kb) from 13ZX28 encodes blaNDM-1. Interestingly, its sequence was identical to the two plasmids p13ZX36–200 (~200-kb) and p13ZX36–70 (~70-kb) from 13ZX36. Formation of the former episome possibly involved homologous recombination through a 4948-bp large fragment located on each of the two latter plasmids. Furthermore, plasmid p13ZX28–272 could be resolved into a ~ 98-kb daughter plasmid by IS26 rearrangement following conjugation. The plasticity of the plasmids is recognized, which warrants further investigation to evaluate the underlying public health risk and understand how antibiotic selection pressure drives this process.</div>","PeriodicalId":49689,"journal":{"name":"Plasmid","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2021-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.plasmid.2021.102568","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25409461","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2

An X1α plasmid from a Salmonella enterica serovar Ohio isolate carrying a novel IS26-bounded tet(C) pseudo-compound transposon 俄亥俄州肠炎沙门氏菌血清分离株携带新型is26结合tet(C)伪复合转座子的X1α质粒

IF 2.6 4区生物学

Plasmid Pub Date : 2021-03-01 DOI: 10.1016/j.plasmid.2021.102561

Carol H. Pong, Ruth M. Hall

{"title":"An X1α plasmid from a Salmonella enterica serovar Ohio isolate carrying a novel IS26-bounded tet(C) pseudo-compound transposon","authors":"Carol H. Pong, Ruth M. Hall","doi":"10.1016/j.plasmid.2021.102561","DOIUrl":"10.1016/j.plasmid.2021.102561","url":null,"abstract":"<div>The sequence of a conjugative plasmid, pSRC22-2, found in a multiply antibiotic resistant Salmonella enterica serovar Ohio isolate SRC22 originally cultured from swine in 1999, was determined. Plasmid pSRC22-2 has a copy number of approximately 40 and transfers tetracycline resistance at very high frequency. It was typed as IncX1 using the three typing schemes proposed for X-type plasmids, which utilize the replication region, iteron region and taxC conjugation gene and pSRC22-2 belongs to the X1α subgroup. The plasmid backbone, derived by removing mobile elements, is shared with pOLA52, which was the first fully sequenced IncX1 plasmid, and five other X1α plasmids. The pSRC22-2 backbone is interrupted by a complete copy of an IS903 isoform, partial copies of IS1 and IS903 on either side of a 5930 bp IS26-bounded pseudo-compound transposon (PCT), and a novel 256 bp miniature inverted repeat transposable element (MITE). The MITE belongs to the Tn3 family and was named MITESen1. The PCT, which carries a tet(C) tetracycline resistance determinant, is bounded by copies of a novel IS26 variant, IS26-v4, and was designated PTn6184. Comparison of PTn6184 with other tet(C)-carrying PCTs revealed that it can be derived from the largest, PTntet(C), via a two-step process that re-orders the central fragment and involves both an IS26-mediated event and homologous recombination. IS26-v4, which encodes a variant transposase, Tnp26 G184D, has appeared in only 46 entries in the GenBank non-redundant database.</div>","PeriodicalId":49689,"journal":{"name":"Plasmid","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2021-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.plasmid.2021.102561","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38772482","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 1

Identification and characterization of a spreadable IncI1 plasmid harbouring a blaCTX-M-15 gene in an Italian human isolate of Salmonella serovar Napoli 意大利那不勒斯人血清沙门氏菌分离株携带blaCTX-M-15基因的传染性IncI1质粒的鉴定和特性

IF 2.6 4区生物学

Plasmid Pub Date : 2021-03-01 DOI: 10.1016/j.plasmid.2021.102566

Sara Petrin , Massimiliano Orsini , Eleonora Mastrorilli , Alessandra Longo , Debora Cozza , John E. Olsen , Antonia Ricci , Carmen Losasso , Lisa Barco

{"title":"Identification and characterization of a spreadable IncI1 plasmid harbouring a blaCTX-M-15 gene in an Italian human isolate of Salmonella serovar Napoli","authors":"Sara Petrin , Massimiliano Orsini , Eleonora Mastrorilli , Alessandra Longo , Debora Cozza , John E. Olsen , Antonia Ricci , Carmen Losasso , Lisa Barco","doi":"10.1016/j.plasmid.2021.102566","DOIUrl":"10.1016/j.plasmid.2021.102566","url":null,"abstract":"<div>Salmonella enterica subsp. enterica serovar Napoli (S. Napoli) ranks among the top serovars causing human infections in Italy, although not common in other European countries. Isolates are generally pan-susceptible or resistant to aminoglycosides only, however data on antimicrobial resistance genes in strains of S. Napoli are limited. Recently an isolate encoding resistance to third generation cephalosporins was reported. This study aimed to characterize plasmid-encoded cephalosporin resistance due to the blaCTX-M-15 gene in a human S. Napoli isolate in Italy, and to investigate plasmid stability over time.S. Napoli 16/174478 was confirmed to be ESBL-producing. The blaCTX-M-15 gene was shown to be located on an IncI1α plasmid of 90,272 bp (50.03 GC%) encoding for 107 coding sequences (CDS). The plasmid was successfully transferred by conjugation to an E. coli 1816 recipient strain (conjugation frequency 3.9 × 10−2 transconjugants per donor). Transconjugants were confirmed to carry the IncI1α plasmid, and to be ESBL-producing strains as well. Moreover, transconjugant colonies maintained the plasmid for up to 10 passages. The identification of S. Napoli isolates able to produce ESBLs is of great concern, as this pathogen is frequently associated with invasive infections and a higher risk of bacteraemia, and its reservoir has not yet been clearly identified.</div>","PeriodicalId":49689,"journal":{"name":"Plasmid","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2021-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.plasmid.2021.102566","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25369513","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 3

Genomic islands related to Salmonella genomic island 1; integrative mobilisable elements in trmE mobilised in trans by A/C plasmids 与沙门氏菌基因组岛1相关的基因组岛;trmE中的整合可移动元件被A/C质粒在trans中动员

IF 2.6 4区生物学

Plasmid Pub Date : 2021-03-01 DOI: 10.1016/j.plasmid.2021.102565

Claire de Curraize , Eliane Siebor , Catherine Neuwirth

{"title":"Genomic islands related to Salmonella genomic island 1; integrative mobilisable elements in trmE mobilised in trans by A/C plasmids","authors":"Claire de Curraize , Eliane Siebor , Catherine Neuwirth","doi":"10.1016/j.plasmid.2021.102565","DOIUrl":"10.1016/j.plasmid.2021.102565","url":null,"abstract":"<div>Salmonella genomic island 1 (SGI1), an integrative mobilisable element (IME), was first reported 20 years ago, in the multidrug resistant Salmonella Typhimurium DT104 clone. Since this first report, many variants and relatives have been found in Salmonella enterica and Proteus mirabilis. Thanks to whole genome sequencing, more and more complete sequences of SGI1-related elements (SGI1-REs) have been reported in these last few years among Gammaproteobacteria. Here, the genetic organisation and main features common to SGI1-REs are summarised to help to classify them. Their integrases belong to the tyrosine-recombinase family and target the 3′-end of the trmE gene. They share the same genetic organisation (integrase and excisionase genes, replicase module, SgaCD-like transcriptional activator genes, traN, traG, mpsB/mpsA genes) and they harbour AcaCD binding sites promoting their excision, replication and mobilisation in presence of A/C plasmid. SGI1-REs are mosaic structures suggesting that recombination events occurred between them. Most of them harbour a multiple antibiotic resistance (MAR) region and the plasticity of their MAR region show that SGI1-REs play a key role in antibiotic resistance and might help multiple antibiotic resistant bacteria to adapt to their environment. This might explain the emergence of clones with SGI1-REs.</div>","PeriodicalId":49689,"journal":{"name":"Plasmid","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2021-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.plasmid.2021.102565","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25369515","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 10

Novel class 1 integron harboring antibiotic resistance genes in wastewater-derived bacteria as revealed by functional metagenomics 功能宏基因组研究发现废水源细菌中含有抗生素耐药基因的新型1类整合子

IF 2.6 4区生物学

Plasmid Pub Date : 2021-03-01 DOI: 10.1016/j.plasmid.2021.102563

Bridget B. McGivern , Rylie K. McDonell , Sydney K. Morris , Timothy M. LaPara , Justin J. Donato

{"title":"Novel class 1 integron harboring antibiotic resistance genes in wastewater-derived bacteria as revealed by functional metagenomics","authors":"Bridget B. McGivern , Rylie K. McDonell , Sydney K. Morris , Timothy M. LaPara , Justin J. Donato","doi":"10.1016/j.plasmid.2021.102563","DOIUrl":"10.1016/j.plasmid.2021.102563","url":null,"abstract":"<div>Combatting antibiotic resistance is critical to our ability to treat infectious diseases. Here, we identified and characterized diverse antimicrobial resistance genes, including potentially mobile elements, from synthetic wastewater treatment microcosms exposed to the antibacterial agent triclosan. After seven weeks of exposure, the microcosms were subjected to functional metagenomic selection across 13 antimicrobials. This was achieved by cloning the combined genetic material from the microcosms, introducing this genetic library into E. coli, and selecting for clones that grew on media supplemented with one of the 13 antimicrobials. We recovered resistant clones capable of growth on media supplemented with a single antimicrobial, yielding 13 clones conferring resistance to at least one antimicrobial agent. Antibiotic susceptibility analysis revealed resistance ranging from 4 to >50 fold more resistant, while one clone showed resistance to multiple antibiotics. Using both Sanger and SMRT sequencing, we identified the predicted active gene(s) on each clone. One clone that conferred resistance to tetracycline contained a gene encoding a novel tetA-type efflux pump that was named TetA(62). Three clones contained predicted active genes on class 1 integrons. One integron had a previously unreported genetic arrangement and was named In1875. This study demonstrated the diversity and potential for spread of resistance genes present in human-impacted environments.</div>","PeriodicalId":49689,"journal":{"name":"Plasmid","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2021-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.plasmid.2021.102563","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25311989","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 3

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