Skip to main content
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems
  • Log in
  • My alerts
  • My Cart

Main menu

  • Home
  • Articles
    • Current Issue
    • Accepted Manuscripts
    • Archive
    • Minireviews
    • JB Special Collection
    • JB Classic Spotlights
  • For Authors
    • Submit a Manuscript
    • Scope
    • Editorial Policy
    • Submission, Review, & Publication Processes
    • Organization and Format
    • Errata, Author Corrections, Retractions
    • Illustrations and Tables
    • Nomenclature
    • Abbreviations and Conventions
    • Publication Fees
    • Ethics Resources and Policies
  • About the Journal
    • About JB
    • Editor in Chief
    • Editorial Board
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
  • Subscribe
    • Members
    • Institutions
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems

User menu

  • Log in
  • My alerts
  • My Cart

Search

  • Advanced search
Journal of Bacteriology
publisher-logosite-logo

Advanced Search

  • Home
  • Articles
    • Current Issue
    • Accepted Manuscripts
    • Archive
    • Minireviews
    • JB Special Collection
    • JB Classic Spotlights
  • For Authors
    • Submit a Manuscript
    • Scope
    • Editorial Policy
    • Submission, Review, & Publication Processes
    • Organization and Format
    • Errata, Author Corrections, Retractions
    • Illustrations and Tables
    • Nomenclature
    • Abbreviations and Conventions
    • Publication Fees
    • Ethics Resources and Policies
  • About the Journal
    • About JB
    • Editor in Chief
    • Editorial Board
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
  • Subscribe
    • Members
    • Institutions
Plant Microbiology

The Twin Arginine Transport System Appears To Be Essential for Viability in Sinorhizobium meliloti

Brad S. Pickering, Ivan J. Oresnik
Brad S. Pickering
Department of Microbiology, University of Manitoba, Winnipeg, Manitoba R3T 2N2, Canada
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Ivan J. Oresnik
Department of Microbiology, University of Manitoba, Winnipeg, Manitoba R3T 2N2, Canada
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: oresniki@cc.umanitoba.ca
DOI: 10.1128/JB.00206-10
  • Article
  • Figures & Data
  • Info & Metrics
  • PDF
Loading

Article Figures & Data

Figures

  • Tables
  • FIG. 1.
    • Open in new tab
    • Download powerpoint
    FIG. 1.

    Genetic region containing tatABC of S. meliloti. A schematic diagram showing the relative positions of the relevant primers used to construct and verify constructs in the tat region. Numbered arrows correspond to the primers listed in Table 2. The plasmid above corresponds to pBP121, whereas the plasmid below corresponds to pBP79. Details of the construction and verification are in the text. The arrow above the Plac promoter indicates the direction of transcription nptII, neomycin phosphotransferase cassette.

  • FIG. 2.
    • Open in new tab
    • Download powerpoint
    FIG. 2.

    Stability of pBP79 containing the tatABC operon. Presence of pBP79 was measured by the proportion of Tc-resistant colonies found at each subculture. Error bars represent standard deviations of three independent experiments. Where not shown, error bars are smaller than the symbol. ▴, SRmA947; ⧫, SRmA938.

  • FIG. 3.
    • Open in new tab
    • Download powerpoint
    FIG. 3.

    Effect of tatABC mutation on cell morphology and viability in S. meliloti. (A and C) Rm1021. (B and D) SRmA947. Panels A and B show DIC images of S. meliloti cells. Panels C and D show cells that were stained using a BacLight LIVE/DEAD bacterial viability kit (Molecular Probes, Invitrogen) and viewed using a fluorescent microscope. Live cells fluoresce green, whereas dead cells fluoresce red. (E) Percentage of live cells in cultures of Rm1021, SRmA938, and SRmA947, stained using a BacLight LIVE/DEAD kit and quantitated using fluorescence. Data are presented as the mean of three independent replicates. Values in parentheses represent standard deviations.

Tables

  • Figures
  • TABLE 1.

    Bacterial strains and plasmids

    Strain or plasmidRelevant characteristic(s)aReference or source
    Strains
        S. meliloti
            Rm1021SU47 str-21 Smr 35
            Rm2011SU47 str-3 Smr 11
            Rm5000SU47 rif-5 Rifr 19
            SRmA363Rm1021 expR+ 38
            SRmA938Rm1021 pKnock-Gm containing 5′ nptII::3′ flanking tatABC This work
            SRmA946Rm1021 pRK7813tatABC::pKnock-GmThis work
            SRmA947Rm1021 tatABCΔ/pRK7813tatABC This work
        S. medicae
            SmD100WSM419 Smr This work
            SmD104SmD100(pBP92)This work
            SmD107SmD107(pBP93)This work
        E. coli
            DH5αλ− φlacZDM15 D(lacZYA-argF)U169 recA1 endA hsdR17(rK− mK −) supE44 thi-1 gyrA relA1 24
            S17-1 recA derivative of MM294A with integrated RP4-2 (Tc::Mu::Km::Tn7) 58
            MM294A pro-82 thi-1 hsdR17 supE44 20
            MT607MM294A recA56 20
            MT616MT607(pRK600) 20
    Plasmids
        pEX18TcBroad-host-range gene replacement vector; sacB Tcr 26
        pBlueScript II SKCloning vector; ColE1 oriV Apr Stratagene
        pRK600pRK2013 npt::Tn9 Cmr 20
        pRK7813RK2 derivative carrying pUC9 polylinker and lambda cos site; Tcr 27
        pKnock-GmSuicide vector for insertional mutagenesis; R6K ori RP4 oriT Gmr 2
        pPH1JIIncP plasmid; Gmr 7
        pBP59 sacB from pEX18Tc inserted into pRK7813This work
        pBP79pBP59 with tatABC cloned into polylinkerThis work
        pBP91pRK7813 tatC::nptII; oriented in the same direction as Plac This work
        pBP92pRK7813 tatC::nptII; oriented in the opposite direction as Plac This work
        pBP103pKnock-Gm containing upstream 5′ tatABC fragmentThis work
        pBP105pBP103containing 3′ downstream tatABC fragmentThis work
        pBP121pBP105 containing nptII cassetteThis work
        pMM4 tatC fragment cloned into pBSThis work
        pMM13Kanr fragment from pMM22 cloned into pMM4This work
        pMM22Kanr fragment cloned as a SmaI fragment into pBlueScriptThis work
    • ↵ a Cm, chloramphenicol; Gm, gentamicin; Kan, kanamycin; Nm, neomycin; Ap, ampicillin; Sm, streptomycin.

  • TABLE 2.

    Primers used in this work

    No.Primer nameSequence (5′ → 3′)
    1sacB FwdATATCCCGGGCCATGGCCATCACATATACCTGCCGTTC
    2sacB RvsATATCCCGGGCCATGGTTATTTGTTAACTGTTAATTGTCCTTG
    3tatABC FwdATATGAATTCTGCGCGAGACGCGCGCGA
    4tatABC RvsATATAAGCTTCTCCTTCGACGCAGCATTGCG
    55′ upstream tat FwdATATGCGGCCGCGGTTTCAGATGCGCGTAACG
    65′ upstream tat RvsATATCCCGGGGTATGGCCAGCGTGTCGGC
    73′ downstream tat FwdATATCCCGGGACCGCGGAGTTCTTGCGCC
    83′ downstream tat RvsATATGGTACCCGACATTTGCGCTTTCGTCCG
    9tatC FwdATATGAATTCCGCCGGCTGCGAATAAG
    10tatC RvsATATGAATTCGCTGCATGACATCGGAG
    11tat confirmation FwdCACCGAAGCCTGAAGAAGAC
    12Tat confirmation RvsGCTCCTTCGAAGTCCATCAG
    13 nptII out LeftTTCGGAATCGTTTTCCGGGAG
    14 nptII out RightTTAGCAGCCCTTGCGCCCTG
  • TABLE 3.

    Frequency of single-crossover recombinants is dependent upon the orientation of Plac a

    ConstructOrientation relative to PlacNo. of recombinants
    S. melilotiS. medicae
    pBP91Same direction1.8 × 10−4 1.7 × 10−4
    pBP92Opposite direction<10−8 <10−8
    • ↵ a Plasmid pBP91 or pBP92 were conjugated from E. coli into either S. meliloti or S. medicae and recombinants were selected on the basis of antibiotic resistance. Data presented are the mean of one experiment with three independent replicates where the standard deviation is less than 10%. The experiment was also replicated on three separate occasions with comparable data.

PreviousNext
Back to top
Download PDF
Citation Tools
The Twin Arginine Transport System Appears To Be Essential for Viability in Sinorhizobium meliloti
Brad S. Pickering, Ivan J. Oresnik
Journal of Bacteriology Sep 2010, 192 (19) 5173-5180; DOI: 10.1128/JB.00206-10

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Print

Alerts
Sign In to Email Alerts with your Email Address
Email

Thank you for sharing this Journal of Bacteriology article.

NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. We do not retain these email addresses.

Enter multiple addresses on separate lines or separate them with commas.
The Twin Arginine Transport System Appears To Be Essential for Viability in Sinorhizobium meliloti
(Your Name) has forwarded a page to you from Journal of Bacteriology
(Your Name) thought you would be interested in this article in Journal of Bacteriology.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
The Twin Arginine Transport System Appears To Be Essential for Viability in Sinorhizobium meliloti
Brad S. Pickering, Ivan J. Oresnik
Journal of Bacteriology Sep 2010, 192 (19) 5173-5180; DOI: 10.1128/JB.00206-10
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Top
  • Article
    • ABSTRACT
    • MATERIALS AND METHODS
    • RESULTS
    • DISCUSSION
    • ACKNOWLEDGMENTS
    • FOOTNOTES
    • REFERENCES
  • Figures & Data
  • Info & Metrics
  • PDF

KEYWORDS

Bacterial Proteins
Biological Transport
Membrane Transport Proteins
Sinorhizobium meliloti

Related Articles

Cited By...

About

  • About JB
  • Editor in Chief
  • Editorial Board
  • Policies
  • For Reviewers
  • For the Media
  • For Librarians
  • For Advertisers
  • Alerts
  • RSS
  • FAQ
  • Permissions
  • Journal Announcements

Authors

  • ASM Author Center
  • Submit a Manuscript
  • Article Types
  • Ethics
  • Contact Us

Follow #Jbacteriology

@ASMicrobiology

       

ASM Journals

ASM journals are the most prominent publications in the field, delivering up-to-date and authoritative coverage of both basic and clinical microbiology.

About ASM | Contact Us | Press Room

 

ASM is a member of

Scientific Society Publisher Alliance

 

American Society for Microbiology
1752 N St. NW
Washington, DC 20036
Phone: (202) 737-3600

Copyright © 2021 American Society for Microbiology | Privacy Policy | Website feedback

Print ISSN: 0021-9193; Online ISSN: 1098-5530