Plasmid transduction using bacteriophage Phi(adh) for expression of CC chemokines by Lactobacillus gasseri ADH.

Item request has been placed! ×
Item request cannot be made. ×
loading   Processing Request
Read More Add to Saved list
  • Author(s): Damelin LH;Damelin LH; Mavri-Damelin D; Klaenhammer TR; Tiemessen CT
  • Source:
    Applied and environmental microbiology [Appl Environ Microbiol] 2010 Jun; Vol. 76 (12), pp. 3878-85. Date of Electronic Publication: 2010 Apr 23.
  • Publication Type:
    Journal Article; Research Support, Non-U.S. Gov't
  • Language:
    English
  • Additional Information
    • Source:
      Publisher: American Society for Microbiology Country of Publication: United States NLM ID: 7605801 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1098-5336 (Electronic) Linking ISSN: 00992240 NLM ISO Abbreviation: Appl Environ Microbiol Subsets: MEDLINE
    • Publication Information:
      Original Publication: Washington, American Society for Microbiology.
    • Subject Terms:
      Genetic Engineering* ; Plasmids* ; Transduction, Genetic*; Bacteriophages/*genetics ; Chemokines, CC/*biosynthesis ; Chemokines, CC/*metabolism ; Lactobacillus/*genetics; Chemokines, CC/genetics ; Humans ; Lactobacillus/metabolism ; Recombinant Proteins/biosynthesis ; Recombinant Proteins/genetics ; Recombinant Proteins/metabolism
    • Abstract:
      Vaginal mucosal microfloras are typically dominated by Gram-positive Lactobacillus species, and colonization of vaginal mucosa by exogenous microbicide-secreting Lactobacillus strains has been proposed as a means of enhancing this natural mucosal barrier against human immunodeficiency virus (HIV) infection. We asked whether an alternative strategy could be utilized whereby anti-HIV molecules are expressed within the cervicovaginal milieu by endogenous vaginal Lactobacillus populations which have been engineered in situ via transduction. In this study, we therefore investigated the feasibility of utilizing transduction for the expression of two HIV coreceptor antagonists, the CC chemokines CCL5 and CCL3, in a predominant vaginal Lactobacillus species, Lactobacillus gasseri. Modifying a previously established transduction model, which utilizes L. gasseri ADH and its prophage Phiadh, we show that mitomycin C induction of L. gasseri ADH transformants containing pGK12-based plasmids with CCL5 and CCL3 expression and secretion cassettes (under the control of promoters P6 and P59, respectively) and a 232-bp Phiadh cos site fragment results in the production of transducing particles which contain 8 to 9 copies of concatemeric plasmid DNA. High-frequency transduction for these particles (almost 6 orders of magnitude greater than that for pGK12 alone) was observed, and transductants were found to contain recircularized expression plasmids upon subsequent culture. Importantly, transductants produced CC chemokines at levels comparable to those produced by electroporation-derived transformants. Our findings therefore lend support to the potential use of transduction in vaginal Lactobacillus species as a novel strategy for the prevention of HIV infection across mucosal membranes.
    • References:
      Am J Reprod Immunol. 2008 Jan;59(1):44-54. (PMID: 18154595)
      J Clin Microbiol. 2002 Aug;40(8):2746-9. (PMID: 12149323)
      J Appl Microbiol. 2007 May;102(5):1337-49. (PMID: 17448169)
      Methods Mol Biol. 1995;47:201-8. (PMID: 7550736)
      J Leukoc Biol. 2003 Sep;74(3):448-55. (PMID: 12949249)
      Plasmid. 1991 Jul;26(1):55-66. (PMID: 1840693)
      Appl Environ Microbiol. 2008 Aug;74(15):4626-35. (PMID: 18539799)
      Appl Environ Microbiol. 2001 Mar;67(3):1253-61. (PMID: 11229918)
      Biochem Pharmacol. 2006 Sep 14;72(6):739-48. (PMID: 16844091)
      Plasmid. 2006 May;55(3):184-93. (PMID: 16458963)
      J Infect Dis. 1999 Dec;180(6):1950-6. (PMID: 10558952)
      J Bacteriol. 1981 Jul;147(1):1-8. (PMID: 6787022)
      Appl Environ Microbiol. 2004 Oct;70(10):6076-85. (PMID: 15466553)
      Appl Environ Microbiol. 1987 Oct;53(10):2452-7. (PMID: 2447829)
      Appl Environ Microbiol. 1984 Oct;48(4):726-31. (PMID: 6095756)
      Appl Environ Microbiol. 1989 Sep;55(9):2206-13. (PMID: 2508554)
      Appl Environ Microbiol. 1992 Jan;58(1):187-93. (PMID: 16348621)
      Nihon Saikingaku Zasshi. 1971 Oct;26(10):482-7. (PMID: 5169937)
      J Acquir Immune Defic Syndr. 2005 Dec 15;40(5):512-20. (PMID: 16284525)
      Appl Environ Microbiol. 1993 Aug;59(8):2730-3. (PMID: 16349028)
      Plasmid. 2001 Sep;46(2):106-16. (PMID: 11591136)
      Methods Mol Med. 1999;17:3-10. (PMID: 21380650)
      Mol Gen Genet. 1990 Sep;223(2):185-91. (PMID: 2123518)
      Can J Microbiol. 1997 Jan;43(1):61-9. (PMID: 9057296)
      AIDS. 2006 Oct 3;20(15):1917-22. (PMID: 16988512)
      Gene. 1993 Apr 15;126(1):61-6. (PMID: 8472961)
      FEBS Lett. 1999 Aug 27;457(2):219-22. (PMID: 10471782)
      J Appl Microbiol. 2002;92(3):451-9. (PMID: 11872120)
      J Immunol. 2006 May 1;176(9):5627-36. (PMID: 16622032)
    • Accession Number:
      0 (Chemokines, CC)
      0 (Recombinant Proteins)
    • Publication Date:
      Date Created: 20100427 Date Completed: 20100928 Latest Revision: 20231105
    • Publication Date:
      20240829
    • Accession Number:
      PMC2893477
    • Accession Number:
      10.1128/AEM.00139-10
    • Accession Number:
      20418431