The
Sullivan Laboratory
Molecular,
Cell, and Developmental Biology
University of California at Santa Cruz
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Testing for Wolbachia infection in C. elegans isolates
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Survey for Wolbachia infection in C.elegans isolates
(Susan Strome and Catharina Casper-Lindley)
We are interested in developing a C.
elegans model for Wolbachia
infections of filarial worms. As a first step toward that goal,
we examined 75 wild isolates of C.
elegans (listed below) to detect strains that are naturally
infected with Wolbachia.
Two screening methods were used, a visual screen and a PCR-based
screen.
1) Visual screen: 15-20 worms from each strain were fixed
in methanol/acetone and stained with the DNA intercalator DAPI to look
for evidence of intracellular bacteria, especially in embryos and the
germ line.
2) PCR screen: DNA was extracted from 10 worms per
strain and PCR performed using Wolbachia-specific primers for 16SrDNA
and ftsZ (sequence from Casiraghi et al. 2001, Parasitology). As
a control for the DNA extraction and the PCR reaction we used
C.elegans-specific primers. PCR cycling conditions: 92˚C for 2
min, 34x (92˚C for 30s, 55˚C for 40s, 72˚C for 2 min), 72˚C for 5 min.
We did not detect Wolbachia
by either assay in any of the 75 C.
elegans wild strains tested. The ftsZ primers were found
to react with E.coli from the
worm culture plates and were disregarded. 16SrDNA primers were
negative in all samples. (Thanks to Theresa Stiernagle and the
Caenorhabditis Genetics Center for providing the C. elegans strains.)
the following strains were used:
AB1
AB2
AB3
AB4
CB3192
CB3193
CB3194
CB3195
CB3196
CB3197
CB3198
CB3199
CB4507
CB4512
CB4555
CB4851
CB4852
CB4853
CB4854
CB4855
CB4856
CB4858
CB4932
CC1
CC2
CC3
DH424
DP13
DR1344
DR1345
DR1346
DR1347
DR1348
DR1349
DR1350
JU258
JU262
JU263
KR314
LSJ1
MY1
MY2
MY5
MY6
MY7
MY8
MY9
MY10
MY11
MY12
MY13
MY14
MY15
MY16
MY17
MY18
MY19
MY20
MY21
MY22
MY23
N2
N2 (ancestral)
PB303
PB306
PX174
PX176
PX178
PX179
RC301
RW6999
RW7000
TR388
TR389
TR403
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Last
updated: May 1, 2006
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