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oai:figshare.com:article/34026912 2026-09-29T17:38:38Z category_4 category_8 category_13 category_14 category_873 category_46 category_734 category_106 category_132 portal_5 item_type_1 month_year_09_2026

<p>Expression of <i>B. burgdorferi mgsA</i> in the RF spirochete <i>B. hermsii</i> and effect on host infectivity.</p> Dan Drecktrah (36180) Laura S. Hall (12206815) Britney Cheff (25138443) Crystal Richards (12206818) Sandra J. Raffel (12240026) Michael Wulf (12206824) Ian T. Bailey (25138446) Frank Gherardini (40103) D. Scott Samuels (12206833) Biochemistry Microbiology Genetics Molecular Biology Chemical Sciences not elsewhere classified Immunology Biological Sciences not elsewhere classified Science Policy Infectious Diseases translation modification system reach high numbers amino acid residues activate carbonyl stress lyme disease spirochete carbonyl stress pathway relapsing fever spirochete div >< p lyme disease spirochetes enigmatic metabolite methylglyoxal burgdorferi mgsa </ carbonyl stress lyme disease relapsing fever mgsa </ burgdorferi </ mayonii </ hermsii </ borreliella </ borrelia </ bb0364 </ significantly reduces results identify relapse phase provide evidence previously believed phosphate imbalance peak level new post necessarily lethal murine infection mouse model metabolic regulation including glycerol dihydroxyacetone phosphate cellular carbohydrate <p>(A) <i>B. hermsii</i> was transformed with <i>B. burgdorferi mgsA</i> under control of its native <i>B. burgdorferi</i> promoter and inserted in lp200. Whole cell extracts of three independent clones, as well as wild-type <i>B. burgdorferi</i> (<i>Bb</i>) and wild-type <i>B. hermsii</i> (<i>Bh</i>), were separated by SDS-PAGE and analyzed by immunoblot with anti-MgsA antibodies to confirm MgsA expression and anti-FlaB antibodies as a loading control. <b>(B)</b> Wild-type <i>B. hermsii</i> (<i>Bh</i>) and <i>B. hermsii</i> expressing the <i>B. burgdorferi mgsA</i> with the gentamicin resistance cassette (<i>Bh</i> + <i>mgsA</i>_Bb) were grown in normal growth medium and cell lysates were either collected (0h) or placed in nutrient stress medium (RPMI) containing 100 µM glycerol (gly 100), 500 µM glycerol (gly 500), neopeptone (neo) or BSK lacking BSA (-BSA). As a positive control for glycation, wild-type <i>B. burgdorferi</i> was incubated in RPMI containing 100 µM glycerol (<i>Bb</i> gly). After 24 h in the nutrient stress media, whole cell lysates were collected, samples were separated by SDS-PAGE and analyzed by immunoblot using antibodies against MG (glycated proteins) or FlaB. <b>(C)</b> Four mice were injected with 1 × 10³ cells of wild-type <i>B. hermsii</i> carrying the gentamicin resistance cassette (<i>Bh</i>+gent^R) or four mice were injected with <i>B. hermsii</i> expressing <i>B. burgdorferi mgsA</i> (<i>Bh</i>+<i>mgsA</i>_Bb). Starting two days post-inoculation (PI), blood was collected daily and spirochetes enumerated by qPCR. The spirochete load over time is plotted for each mouse using a different color. <b>(D)</b> The levels (log cells/ml blood) of peak spirochetemia was plotted for each mouse infected with either <i>Bh</i>+gent^R or <i>Bh</i>+<i>mgsA</i>_Bb. Bars represent the mean and asterisk represents <i>P</i> = 0.04 as determined by Student’s <i>t</i>-test.</p> 2026-09-29T17:55:00Z Image Figure 10.1371/journal.ppat.1014657.g011 https://figshare.com/articles/figure/_p_Expression_of_i_B_burgdorferi_mgsA_i_in_the_RF_spirochete_i_B_hermsii_i_and_effect_on_host_infectivity_p_/34026912 CC BY 4.0