IF 2.6 4区生物学

Plasmid Pub Date : 2020-07-01 DOI: 10.1016/j.plasmid.2020.102513

Özge Uğurlu , Fırat Barış Barlas , Serap Evran , Suna Timur

{"title":"The cell-penetrating YopM protein-functionalized quantum dot-plasmid DNA conjugate as a novel gene delivery vector","authors":"Özge Uğurlu , Fırat Barış Barlas , Serap Evran , Suna Timur","doi":"10.1016/j.plasmid.2020.102513","DOIUrl":"10.1016/j.plasmid.2020.102513","url":null,"abstract":"<div>Non-viral gene delivery systems have great potential for safe and efficient gene therapy, while inefficient cellular and nuclear uptake remain as the major hurdles. Novel approaches are needed to enhance the transfection efficiency of non-viral vectors. In accordance with this need, the objective of this study was to construct a non-viral vector that could achieve gene delivery without using additional lipid-based transfection agent. We aimed to impart self-delivery property to a non-viral vector by using the cell and nucleus penetrating properties of YopM proteins from the three Yersinia spp. (Y. pestis, Y. enterocolotica and Y. pseudotuberculosis). Plasmid DNA (pDNA) encoding green fluorescent protein (GFP) was labeled with quantum dots (QDs) via peptide-nucleic acid (PNA) recognition site. Recombinant YopM protein was then attached to the conjugate via a second PNA recognition site. The YopM ̶ QDs ̶ pDNA conjugate was transfected into HeLa cells without using additional transfection reagent. All three conjugates produced GFP fluorescence, indicating that the plasmid was successfully delivered to the nucleus. As control, naked pDNA was transfected into the cells by using a commercial transfection reagent. The Y. pseudotuberculosis YopM-functionalized conjugate achieved the highest GFP expression, compared to other two YopM proteins and the transfection reagent. To the best of our knowledge, YopM protein was used for the first time in a non-viral gene delivery vector.</div>","PeriodicalId":49689,"journal":{"name":"Plasmid","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2020-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.plasmid.2020.102513","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38013236","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: 7

Construction of a novel CRISPRi-based tool for silencing of multiple genes in Mycobacterium tuberculosis 构建一种基于crispr的新型结核分枝杆菌多基因沉默工具

IF 2.6 4区生物学

Plasmid Pub Date : 2020-07-01 DOI: 10.1016/j.plasmid.2020.102515

Nisheeth Agarwal

{"title":"Construction of a novel CRISPRi-based tool for silencing of multiple genes in Mycobacterium tuberculosis","authors":"Nisheeth Agarwal","doi":"10.1016/j.plasmid.2020.102515","DOIUrl":"10.1016/j.plasmid.2020.102515","url":null,"abstract":"<div>Due to lipid-rich cell wall, slow growth and pathogenic nature, it is difficult to manipulate Mycobacterium tuberculosis (Mtb) genome by conventional tools. Recently we have introduced a novel CRISPRi approach for repression of genes in mycobacteria. Although the existing CRISPRi plasmid is proven useful for silencing individual targets, disruption of multiple ORFs remains challenging in mycobacteria. Herein, we report construction of the guide sequence expressing plasmid, pGrna to facilitate cloning and expression of multiple guide sequence cassettes targeting a versatile set of Mtb genes from a single plasmid. Using the modified plasmid, pGrna2, it was shown that expression of all the 10 extracellular sigma factor-encoding genes together with sigB and sigF can be efficiently repressed in Mtb expressing dCas9. In vitro growth analysis indicates that simultaneous knockdown of these non-essential transcriptional regulators is lethal for growth. Importantly, the Δ12sig strain exhibits sensitivity to transcriptional inhibitor rifampicin and oxidative stress diamide, further implying involvement of these genes in controlling bacterial stress response. To the best of my knowledge, this is the first report wherein 12 genes have been efficiently silenced together in a single recombinant strain of Mtb. The modified pGrna2 plasmid offers a powerful tool to decipher the functioning of genes that are redundant or regulate a particular metabolic pathway and can be useful in identification of novel anti-tuberculosis drug targets.</div>","PeriodicalId":49689,"journal":{"name":"Plasmid","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2020-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.plasmid.2020.102515","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38041995","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: 9

Incompatibility and phylogenetic relationship of I-complex plasmids i -复合体质粒的不亲和性及其系统发育关系

IF 2.6 4区生物学

Plasmid Pub Date : 2020-05-01 DOI: 10.1016/j.plasmid.2020.102502

Marta Rozwandowicz , Joost Hordijk , Alex Bossers , Aldert L. Zomer , Jaap A. Wagenaar , Dik J. Mevius , Michael S.M. Brouwer

{"title":"Incompatibility and phylogenetic relationship of I-complex plasmids","authors":"Marta Rozwandowicz , Joost Hordijk , Alex Bossers , Aldert L. Zomer , Jaap A. Wagenaar , Dik J. Mevius , Michael S.M. Brouwer","doi":"10.1016/j.plasmid.2020.102502","DOIUrl":"10.1016/j.plasmid.2020.102502","url":null,"abstract":"<div>Plasmid incompatibility is the inability of two plasmids to be stably maintained in one cell, resulting in loss of one of the plasmids in daughter cells. Dislodgement is a phenotypically distinct form of incompatibility, described as an imperfect reproduction, manifesting in rapid exclusion of a resident plasmid after superinfection. The relationship between plasmids of the phenotypic incompatibility groups IncB/O and IncZ is unclear. Their inability to co-exist was initially referred to as dislodgement while other research reached the conclusion that IncB/O and IncZ plasmids are incompatible. In this manuscript we re-evaluated the relationship between IncB/O and IncZ plasmids to settle these conflicting conclusions. We performed dislodgement testing of R16Δ (IncB/O) and pSFE-059 (IncZ) plasmids by electroporation in a bacterial cell and checked their stability. Stability tests of the obtained plasmid pair showed that the IncB/O plasmid was exclusively and almost completely lost from the heteroplasmid Escherichia coli population. Other IncB/O – IncZ pairs could not form a heteroplasmid population, using conjugation or electroporation. Our data supports the previous suggestion that IncB/O and IncZ plasmids may be considered phenotypically incompatible.</div>","PeriodicalId":49689,"journal":{"name":"Plasmid","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2020-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.plasmid.2020.102502","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"37738324","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 4

Tailor-made sRNAs: a plasmid tool to control the expression of target mRNAs in Pseudomonas putida 定制的sRNAs:一种控制恶臭假单胞菌靶mrna表达的质粒工具

IF 2.6 4区生物学

Plasmid Pub Date : 2020-05-01 DOI: 10.1016/j.plasmid.2020.102503

Patrícia Apura , Margarida Saramago , Alexandra Peregrina , Sandra C. Viegas , Sandra M. Carvalho , Lígia M. Saraiva , Cecília M. Arraiano , Susana Domingues

{"title":"Tailor-made sRNAs: a plasmid tool to control the expression of target mRNAs in Pseudomonas putida","authors":"Patrícia Apura , Margarida Saramago , Alexandra Peregrina , Sandra C. Viegas , Sandra M. Carvalho , Lígia M. Saraiva , Cecília M. Arraiano , Susana Domingues","doi":"10.1016/j.plasmid.2020.102503","DOIUrl":"10.1016/j.plasmid.2020.102503","url":null,"abstract":"<div>Pseudomonas putida is a highly attractive production system for industrial needs. However, for its improvement as a biocatalyst at the industrial level, modulation of its gene expression is urgently needed. We report the construction of a plasmid expressing a small RNA-based system with the potential to be used for different purposes. Due to the small RNAs modular composition, the design facilities and ability to tune gene expression, they constitute a powerful tool in genetic and metabolic engineering. In the tool presented here, customized sRNAs are expressed from a plasmid and specifically directed to any region of a chosen target. Expression of these customized sRNAs is shown to differentially modulate the level of endogenous and heterologous reporter genes. The antisense interaction of the sRNA with the mRNA produces different outcomes. Depending on the particularity of each sRNA-target mRNA pair, we demonstrate the duality of this system, which is able either to decrease or increase the expression of the same given gene. This system combines high specificity with the potential to be widely applied, due to its predicted ability to modulate the expression of virtually any given gene. This plasmid can be used to redesign P. putida metabolism, fulfilling an important industrial gap.</div>","PeriodicalId":49689,"journal":{"name":"Plasmid","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2020-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.plasmid.2020.102503","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"37768411","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: 14

Strong synthetic stationary phase promoter-based gene expression system for Escherichia coli 基于强合成固定相启动子的大肠杆菌基因表达体系

IF 2.6 4区生物学

Plasmid Pub Date : 2020-05-01 DOI: 10.1016/j.plasmid.2020.102491

Jananee Jaishankar, Preeti Srivastava

{"title":"Strong synthetic stationary phase promoter-based gene expression system for Escherichia coli","authors":"Jananee Jaishankar, Preeti Srivastava","doi":"10.1016/j.plasmid.2020.102491","DOIUrl":"10.1016/j.plasmid.2020.102491","url":null,"abstract":"<div>The Gram-negative bacterium Escherichia coli has been the work horse for recombinant protein production since the past several years. However, most of the gene expression systems used either require expensive inducers or exhibit low strength. In the present study, we have generated a strong promoter by repeated rounds of random mutagenesis in a stationary phase promoter isolated from Gordonia sp. IITR100. The promoter activity increased 16-fold as compared to the wild-type promoter. The resultant synthetic promoter showed β-galactosidase activities of ~16,000 Miller units which is comparable to the strong T7 promoter ~13,000 Miller units. The amount of LacZ produced by the synthetic promoter was found to be active for several days in stationary phase. The advantage of this synthetic promoter over T7 promoter includes its stationary phase auto-inducibility thereby saving the cost of addition of inducers. Expression of GFPuv was observed in all the cells of E. coli due to the absence of requirement of inducer. A general-purpose vector containing the synthetic promoter with an MCS ready for use has been developed in the study. It has also been used to demonstrate the production of two heterologous proteins.</div>","PeriodicalId":49689,"journal":{"name":"Plasmid","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2020-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.plasmid.2020.102491","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"37623061","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

The role of hosts, plasmids and environment in determining plasmid transfer rates: A meta-analysis 宿主、质粒和环境在决定质粒转移率中的作用:一项荟萃分析

IF 2.6 4区生物学

Plasmid Pub Date : 2020-03-01 DOI: 10.1016/j.plasmid.2020.102489

Richard J. Sheppard, Alice E. Beddis, Timothy G. Barraclough

引用次数: 34

ORF-based binarized structure network analysis of plasmids (OSNAp), a novel approach to core gene-independent plasmid phylogeny 基于orf的质粒二值化结构网络分析(opsnap)是一种研究核心基因无关质粒系统发育的新方法

IF 2.6 4区生物学

Plasmid Pub Date : 2020-03-01 DOI: 10.1016/j.plasmid.2019.102477

Masahiro Suzuki , Yohei Doi , Yoshichika Arakawa

{"title":"ORF-based binarized structure network analysis of plasmids (OSNAp), a novel approach to core gene-independent plasmid phylogeny","authors":"Masahiro Suzuki , Yohei Doi , Yoshichika Arakawa","doi":"10.1016/j.plasmid.2019.102477","DOIUrl":"10.1016/j.plasmid.2019.102477","url":null,"abstract":"<div><h3>Objectives</h3>Systematic comparison of multiple plasmids remains challenging. We aimed to develop a new method for phylogenetic analysis of plasmids, open reading frame (ORF)-based binarized structure network analysis of plasmids (OSNAp).</div><div><h3>Methods</h3>With the OSNAp, the genetic structures of plasmids in a given plasmid group are expressed as binary sequences based on the presence or absence of ORFs regardless of their positions or directions. As a proof-of-concept, ORFs were collected from 101 complete I1 plasmid sequences, and their corresponding binary sequences were generated. A tree was generated using the neighbor-net, an algorithm for constructing phylogenetic networks based on distance between taxa, to visualize the plasmid phylogeny drawn from binary sequences. The results were compared with those of plasmid sequence types (pSTs) defined by plasmid multilocus sequence typing (pMLST).</div><div><h3>Results</h3>All I1 plasmids were placed on the phylogenetic tree constructed from the binary sequences. Most plasmids belonging to the same pSTs had Dice indices of ≥0.95 and were placed in the same OSNAp split. On the other hand, pST12 plasmids were distributed on separate splits due to differences in ORFs not used in pMLST, suggesting improved differentiation of the plasmids with OSNAp compared with pMLST.</div><div><h3>Conclusion</h3>OSNAp is a novel holistic approach to assess relatedness of a population of plasmids in a given plasmid group based on nucleotide sequence data. It provides higher discrimination than pMLST, which may prove useful in tracing bacteria that harbor plasmids of shared origins.</div>","PeriodicalId":49689,"journal":{"name":"Plasmid","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2020-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.plasmid.2019.102477","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"37485907","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: 7

Cloning of pAhX22, a small cryptic plasmid from Aeromonas hydrophila, and construction of a pAhX22-derived shuttle vector 嗜水气单胞菌小隐质粒pAhX22的克隆及pAhX22穿梭载体的构建

IF 2.6 4区生物学

Plasmid Pub Date : 2020-03-01 DOI: 10.1016/j.plasmid.2020.102490

Xingyu Kang , Chunqiu Li , Yi Luo

{"title":"Cloning of pAhX22, a small cryptic plasmid from Aeromonas hydrophila, and construction of a pAhX22-derived shuttle vector","authors":"Xingyu Kang , Chunqiu Li , Yi Luo","doi":"10.1016/j.plasmid.2020.102490","DOIUrl":"10.1016/j.plasmid.2020.102490","url":null,"abstract":"<div>In this study, a cryptic plasmid from Aeromonas hydrophila (pAhX22) was cloned and characterized. pAhX22 was 2523 bp long, had a GC content of 59.9%, and contained two putative open reading frames (ORFs). orf1 and orf2 encoded putative proteins of 458 amino acids and 88 amino acids, respectively; these putative proteins might be involved in plasmid replication. An Escherichia coli–A. hydrophila shuttle vector, pAEsv-1 (4587 bp, KanR), was constructed using in-fusion cloning, combining pAhX22 with the kanamycin-resistance gene and the origin of replication from E. coli expression vector pET-28a. The transformation efficiency of pAEsv-1 in A. hydrophila strains ranged from 2.2 × 106 to 1.0 × 107 CFU/μg DNA, while transformation efficiency in E. coli DH5α was about 1.6 × 106 CFU/μg DNA. pAEsv-1 was segregationally and structurally stable in A. hydrophila in the absence of selective pressure. A green fluorescent protein gene (gfp) from pHT315-gfp was successfully cloned and expressed in A. hydrophila strain X2 using pAEsv-1, and 82.3% ± 2.5% of cells maintained the recombinant plasmid after one week in liquid culture without kanamycin. These results suggested that pAEsv-1 might potentially be used as a stable cloning vector for A. hydrophila, which might facilitate genetic studies of A. hydrophila.</div>","PeriodicalId":49689,"journal":{"name":"Plasmid","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2020-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.plasmid.2020.102490","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"37598326","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

SGI0, a relative of Salmonella genomic islands SGI1 and SGI2, lacking a class 1 integron, found in Proteus mirabilis SGI0是沙门氏菌基因组岛SGI1和SGI2的近亲，缺乏1类整合子，发现于变形杆菌中

IF 2.6 4区生物学

Plasmid Pub Date : 2020-01-01 DOI: 10.1016/j.plasmid.2019.102453

Claire de Curraize , Eliane Siebor , Catherine Neuwirth , Ruth M. Hall

{"title":"SGI0, a relative of Salmonella genomic islands SGI1 and SGI2, lacking a class 1 integron, found in Proteus mirabilis","authors":"Claire de Curraize , Eliane Siebor , Catherine Neuwirth , Ruth M. Hall","doi":"10.1016/j.plasmid.2019.102453","DOIUrl":"10.1016/j.plasmid.2019.102453","url":null,"abstract":"<div>Several groups of integrative mobilizable elements (IMEs) that harbour a class 1 integron carrying antibiotic resistance genes have been found at the 3′-end of the chromosomal trmE gene. Here, a new IME, designated SGI0, was found in trmE in the sequenced and assembled genome of a French clinical, multiply antibiotic resistant Proteus mirabilis strain, Pm1LENAR. SGI0 shares the same gene content as the backbones of SGI1 and SGI2 (overall 97.6% and 97.7% nucleotide identity, respectively) but it lacks a class 1 integron. However, SGI0 is a mosaic made up of segments with >98.5% identity to SGI1 and SGI2 interspersed with segments sharing 74–95% identity indicating that further diverged backbone types exist and that recombination between them is occurring. The structure of SGI1-V, here re-named SGI-V, which lacks two SGI1 (S023 and S024) backbone genes and includes a group of additional genes in the backbone, was re-examined. In regions shared with SGI1, the backbones shared 97.3% overall identity with the differences distributed in patches with various levels of identity. The class 1 integron is also in a slightly different position with the target site duplication AAATT instead of ACTTG for SGI1 and variants, indicating that it was acquired independently. The Pm1LENAR resistance genes are in the chromosome, in Tn7 and an ISEcp1-mobilised segment.</div>","PeriodicalId":49689,"journal":{"name":"Plasmid","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2020-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.plasmid.2019.102453","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45699341","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

The Tcp plasmids of Clostridium perfringens require the resP gene to ensure stable inheritance 产气荚膜梭菌的Tcp质粒需要有resP基因才能保证稳定的遗传

IF 2.6 4区生物学

Plasmid Pub Date : 2020-01-01 DOI: 10.1016/j.plasmid.2019.102461

Sarah Revitt-Mills , Carmen Lao , Marie Archambault , Dena Lyras , Julian I. Rood , Vicki Adams

{"title":"The Tcp plasmids of Clostridium perfringens require the resP gene to ensure stable inheritance","authors":"Sarah Revitt-Mills , Carmen Lao , Marie Archambault , Dena Lyras , Julian I. Rood , Vicki Adams","doi":"10.1016/j.plasmid.2019.102461","DOIUrl":"10.1016/j.plasmid.2019.102461","url":null,"abstract":"<div>Many of the disease-causing toxins of the pathogenic bacterium Clostridium perfringens are harboured on large, highly stable, conjugative plasmids. Previous work has established the requirement of a ParMRC-like partitioning system for plasmid maintenance, but little is known about other mechanisms used to ensure stable plasmid inheritance. The archetypal 47 kb Tcp plasmid, pCW3, encodes a gene, resP, whose putative product has sequence similarity to members of the serine recombinase family of site-specific recombinases. ResP is therefore likely to function to resolve plasmid multimers. Sequence analysis identified that resP genes are present on all C. perfringens plasmid families, suggesting a conserved function in these plasmids. To assess the requirement of resP for the stability of pCW3, deletion mutants were constructed. Deletion of resP from pCW3 resulted in a marked instability phenotype that was rescued upon complementation with the wild-type resP gene. Complementation with resP genes from two different C. perfringens plasmids demonstrated that only closely related resP genes can complement the mutation on pCW3. The function of ResP in vivo was examined using an Escherichia coli model system, which determined that two directly repeated res sites were required for the resolution of DNA and that ResP could resolve multimeric plasmid forms into monomeric units. Based on these findings we concluded that ResP could catalyse the resolution of plasmid multimers and was required for the maintenance of Tcp plasmids within C. perfringens. Overall, the results of this study have significant implications for our understanding of the maintenance of toxin-encoding plasmids within C. perfringens.</div>","PeriodicalId":49689,"journal":{"name":"Plasmid","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2020-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.plasmid.2019.102461","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42661560","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

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