<p>Expression of <i>B. burgdorferi mgsA</i> in the RF spirochete <i>B. hermsii</i> and effect on host infectivity.</p>

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).