OspC facilitates Borrelia burgdorferi invasion of Ixodes scapularis salivary glands
J. Clin. Invest. Utpal Pal, et al. 113:220 doi:10.1172/JCI19894 [
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Figure 4Measurement of viable
B. burgdorferi and
ospC gene expression in feeding ticks exposed to OspC F(ab)
2. (
a) Assessment of viable
B. burgdorferi as measured by quantitative PCR of
flaB mRNA in feeding tick gut.
B. burgdorferi–infected nymphs were treated with NRS-F(ab)
2 or OspC-F(ab)
2 (NRS-Fab or OspC-Fab, respectively) were analyzed at 48 or 96 hours after the onset of feeding. Equal amounts of cDNA, converted from total RNA from each group of ticks, were subjected to quantitative PCR. Known quantities of
B. burgdorferi DNA and pCR 2.1 plasmid carrying the tick
β-actin gene were used to prepare standard PCR curves. Amounts of tick
β-actin were determined in each samples and use to normalize the quantities of spirochete DNA between the samples. Differences in
flaB RNA amounts between NRS-Fab and OspC-Fab groups of tick at 48 or 96 hours were not significant (
n = 3). (
b) Measurement of
ospC transcripts in feeding tick gut by quantitative PCR as described above. NRS-Fab and OspC-Fab nymphs were analyzed at 48 or 96 hours after the onset of the feeding. The amounts of
flaB and
ospC transcripts were measured in each samples and data shown are the quantities of
ospC cDNA relative to
flaB cDNA in normalized tick samples. Differences in
ospC cDNA amounts between NRS-Fab and OspC-Fab groups of tick at 48 or 96 hours were not significant (
n = 3).
