• Title/Summary/Keyword: broad host range

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Host Range of pTi12 Contained Agrobacterium tumefaciens KU12 Isolated from Korea (pTi-12를 함유한 한국산 Agrobacterium tumefaciens KU12의 숙주범위)

  • 전경아
    • Journal of Plant Biology
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    • v.33 no.2
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    • pp.97-104
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    • 1990
  • In order to investigate the host range of Agrobacterium tumefaciens KU12 containing pTi-12, 28 species of dicotyledonous plants were infected with KU12, A136 without Ti plasmid and A348 containing pTi A6, respectively. KU12 and A348 induced tumor in 20 species and 14 species, respectively. This results showed that KU12 has a wide host range. Therefore, it was confirmed that KU12 and pTi-12 are very useful for developing plant vector system having a broad host range.

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Properties of Recombinant Derivatives of pJY501, A Multi-copy Streptomyces plasmid (Multi-copy Streptomyces 플라스미드, pJY501의 재조합 유도체의 특성)

  • 염도영;공인수;유주현
    • Microbiology and Biotechnology Letters
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    • v.18 no.1
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    • pp.94-97
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    • 1990
  • The restriction cleavage map of multi-copy recombinant plasmid, pJY502 (5.5 kb), carrying the thiostrepton resistance gene (tsr) was determined. Comparison of the restriction pattern with that of Streptomyces plasmids previously demonstrated that pJY502 was novel. The plasmid pJY502 had a broad host range in Streptomyces and contained single BgtII site for cloning purpose. Transformation frequency of pJY502 was $2.2 \times 10^5$ in S. lividans. E. coti-Streptomyces bifunctional plasmid, pJY504, was also constructed.

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Virulence Factors and Stability of Coliphages Specific to Escherichia coli O157:H7 and to Various E. coli Infection

  • Kim, Eun-Jin;Chang, Hyun-Joo;Kwak, Soojin;Park, Jong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.26 no.12
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    • pp.2060-2065
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    • 2016
  • Characteristics of E. coli O157:H7-specific infection bacteriophages (O157 coliphages) and broad-host-range bacteriophages for other E. coli serotypes (broad-host coliphages) were compared. The burst sizes of the two groups ranged from 40 to 176 PFU/infected cell. Distributions of the virulence factors stx1, stx2, ehxA, and saa between the two groups were not differentiated. Broad-host-range coliphages showed lower stability at $70^{\circ}C$, in relation to O157 coliphages. However, O157 coliphages showed high acid and ethanol tolerance by reduction of only 22% and 11% phages, respectively, under pH 3 and 70% ethanol for 1 h exposure. Therefore, these results revealed that the O157 coliphages might be more stable under harsh environments, which might explain their effective infection of the acid-tolerant E. coli O157:H7.

A Series of IncQ-Based Reporter Plasmids for Use in a Range of Gram-Negative Genera

  • O'Sullivan, Laura E.;Nickerson, Cheryl A.;Wilson, James W.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.5
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    • pp.871-874
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    • 2010
  • Many studies require expression analysis of the same gene/promoter across a range of bacterial genera. However, there is currently a lack of availability of reporters based on the broad-host-range IncQ replicon, which is compatible with a popular improved IncP transfer system that is self-transfer defective. We report IncQ lacZ reporter plasmids with features including (1) compatibility with IncP, IncW, and pBHR/pBBR replicons, (2) a variety of antibiotic markers (Sp-r, Sm-r, Km-r, Cm-r), (3) convenient mobilization via a novel self-transfer-defective IncP conjugation system, and (4) GenBank DNA sequences. Utility is demonstrated using three different promoters in different Gram-negative genera.

Characterization of Recombinant Derivatives of pJY711 of Multicopy Streptomyces Plasmid (Multicopy Streptomyces 플라스미드 pJY711의 재조합 유도체의 특성)

  • 염도영;공인수;유주현
    • Korean Journal of Microbiology
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    • v.28 no.1
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    • pp.35-40
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    • 1990
  • The restriction clevage map of multi-copy recombinant plasmid, pJY712(8.1kb), carrying the thiostrepton resistance gene(tsr) was determined. pJY712 had a broad host range in Streptomyces and contained single BglII site for cloning purpose. The plasmid showed the phenomenon of lethal zygosis ($Ltz^{+}$). Transformation frequency of pJY712 was $5.0\times 10^{4}$ transformants per ug plasmid DNA (TFU) in S. lividans. Plasmid pJY713 was constructed by inserting the tyrosinase gene(mel) into the BclI site of pJY712. Recombinant plasmid pJY714 carrying the mel gene was constructed by in vitro deletion of a segment (1.9kb BglII-BclI fragment) from pJY713.

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Molecular Mechanism of R1162 Plasmid Incompatibility Exerted by Direct Repeat in the Replicative Origin

  • Kim, Yung-Jin
    • BMB Reports
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    • v.29 no.1
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    • pp.63-67
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    • 1996
  • In order to elucidate the molecular mechanism of plasmid incompatibility of broad host-range plasmid R1162, the plasmid-encoded replication protein RepIB was purified and tested for binding to the 20 bp direct repeat (DR) DNA sequence which is reiterated 3 and 1/2 times within the replicative origin of the plasmid. The RepIB protein specifically binds to the DR DNA. Point mutations in the DR which affect expression of plasmid incompatibility also coordinately affect binding. These results indicate that the incompatibility of broad host-range plasmid R1162 is exerted by the DR DNA by titrating the essential replication protein RepIB.

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Studies on Synonymous Codon and Amino Acid Usage Biases in the Broad-Host Range Bacteriophage KVP40

  • Sau Keya;Gupta Sanjib Kumar;Sau Subrata;Mandal Subhas Chandra;Ghosh Tapash Chandra
    • Journal of Microbiology
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    • v.45 no.1
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    • pp.58-63
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    • 2007
  • In this study, the relative synonymous codon and amino acid usage biases of the broad-host range phage, KVP40, were investigated in an attempt to understand the structure and function of its proteins/protein-coding genes, as well as the role of its tRNAs. Synonymous codons in KVP40 were determined to be AT-rich at the third codon positions, and their variations are dictated principally by both mutational bias and translational selection. Further analysis revealed that the RSCU of KVP40 is distinct from that of its Vibrio hosts, V. cholerae and V. parahaemolyticus. Interestingly, the expression of the putative highly expressed genes of KVP40 appear to be preferentially influenced by the abundant host tRNA species, whereas the tRNAs expressed by KVP40 may be required for the efficient synthesis of all its proteins in a diverse array of hosts. The data generated in this study also revealed that KVP40 proteins are rich in low molecular weight amino acid residues, and that these variations are influenced primarily by hydropathy, mean molecular weight, aromaticity, and cysteine content.

Self-Transmissible IncP R995 Plasmids with Alternative Markers and Utility for Flp/FRT Cloning Strategies

  • Santiago, Clayton P.;Quick, Laura N.;Wilson, James W.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.11
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    • pp.1123-1126
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    • 2011
  • The IncP plasmid R995 has been a useful self-transmissible, broad-host-range vector for a number of applications including the recombinase/conjugation-based cloning of large genomic DNA segments. However, R995 derivatives (or related plasmids) expressing a wide range of different resistance markers and Flp recombinase target sites do not exist in the literature. In addition, documented strategies for applying such plasmids in cloning applications that take advantage of conjugation for the convenient isolation and recovery of constructs are extremely limited. Here, we report a new series of R995 plasmids with alternative markers to increase options for applications in backgrounds already expressing resistance to a particular antibiotic(s). These R995 plasmids have been engineered to contain FRT sites that can be used for recombinase-based cloning. We demonstrate the utility of this approach by cloning 20 kb regions from the Salmonella Typhimurium and Escherichia coli genomes and by cloning DNA from an exogenous plasmid source. To our knowledge, this represents the first systematic engineering of an intact, self-transmissible IncP plasmid with a series of alternative antibiotic markers and FRT sites.

Host Cell-Intrinsic Antiviral Defense Induced by Type I Interferons

  • Asano, Atsushi
    • Korean Journal of Agricultural Science
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    • v.35 no.2
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    • pp.177-182
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    • 2008
  • Type I Interferons (IFNs) are potent antiviral cytokines that modulate both innate immunity and adaptive immunity. Type I IFNs are immediately induced by viral infection, and stimulate production of a broad range of gene products such as double-stranded RNA-activated protein kinase (PKR), 2' 5'-oligoadenylate synthetase (OAS)/RNaseL and Mx GTPases. These proteins inhibit viral replication in host cells. Type I IFNs, in turn, lead to antiviral state at early phase of viral infection. We provide an overview of the knowledge of IFN-inducible antiviral proteins conserved in vertebrates.

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A Broad-Host-Range Promoter-Probe Vector, pKU20, and Its Use in Promoter Cloning and Expression of Bacillus thuringiensis Crystal Protein Gene in Pseudomonas putida

  • SHIN, BYUNG SIK;BON TAG KOO;SEUNG HWAN PARK;HO YONG PARK;JEONG IL KIM
    • Journal of Microbiology and Biotechnology
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    • v.1 no.4
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    • pp.240-245
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    • 1991
  • We have constructed a promoter-probe vector pKU20 using pKT230, a derivative of broad-host-range plsmid RSF1010, as a base. The pKU20 contains structural gene for aminoglycoside phos-photransferase (aph), without promoter, and a multiple cloning site upstream the aph. Using this vector, a 412base pairs (bp) PstI fragment showing strong promoter activity both in Escherichia coli LE392 and Pseudomonas putida KCTC1644 has been cloned from Pseudomonas fluorescens chromosomal DNA on the basis of streptomycin resistance. The nucleotide sequence of the 412 bp fragment has been determined and the putative - 35 and -10 region was observed. Insecticidal protein gene of Bacillus thuringiensis subsp. kurstaki HD-73 inserted on downstream of the promoterlike DNA fragment was efficiently expressed in E. coli and P. putida. The toxin protein was efficiently synthesized in an insoluble form in both strains.

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