TY - JOUR
T1 - Culture and Phenotypic Characterization of a Wolbachia pipientis Isolate
AU - Fenollar, Florence
AU - La Scola, Bernard
AU - Inokuma, Hisashi
AU - Dumler, J. Stephen
AU - Taylor, Mark J.
AU - Raoult, Didier
PY - 2003/12
Y1 - 2003/12
N2 - The recent isolation of Wolbachia pipientis in the continuous cell line Aa23, established from eggs of a strain of the Asian tiger mosquito Aedes albopictus, allowed us to perform extensive characterization of the isolate. Bacterial growth could be obtained in C6/36, another A. albopictus cell line, at 28°C and in a human embryonic lung fibroblast monolayer at 28 and 37°C, confirming that its host cell range is broader than was initially thought. The bacteria were best visualized by Diff-Quik and May-Grü nwald-Giemsa staining. Proteins from 213 to 18 kDa with two major protein bands of 65 and 25 kDa were observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. By Western blotting with specific polyclonal mouse and rabbit antisera, dominant immunoreactive antigens were found at approximately 100, 80, and 30 kDa. The genome size was calculated to be 1,790 ± 17 kb by pulsed-field gel electrophoresis. The sequence of the citrate synthase gene (glt,4) of W. pipientis was determined by gene walking. Its position in the phylogenetic tree constructed with gita confirmed that found in a phylogenetic tree constructed with 16S rRNA genes and that it belongs in the α subgroup of the class Proteobacteria and that it is closely related to but independent from the genera Ehrlichia, Anaplasma, and Neorickettsia.
AB - The recent isolation of Wolbachia pipientis in the continuous cell line Aa23, established from eggs of a strain of the Asian tiger mosquito Aedes albopictus, allowed us to perform extensive characterization of the isolate. Bacterial growth could be obtained in C6/36, another A. albopictus cell line, at 28°C and in a human embryonic lung fibroblast monolayer at 28 and 37°C, confirming that its host cell range is broader than was initially thought. The bacteria were best visualized by Diff-Quik and May-Grü nwald-Giemsa staining. Proteins from 213 to 18 kDa with two major protein bands of 65 and 25 kDa were observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. By Western blotting with specific polyclonal mouse and rabbit antisera, dominant immunoreactive antigens were found at approximately 100, 80, and 30 kDa. The genome size was calculated to be 1,790 ± 17 kb by pulsed-field gel electrophoresis. The sequence of the citrate synthase gene (glt,4) of W. pipientis was determined by gene walking. Its position in the phylogenetic tree constructed with gita confirmed that found in a phylogenetic tree constructed with 16S rRNA genes and that it belongs in the α subgroup of the class Proteobacteria and that it is closely related to but independent from the genera Ehrlichia, Anaplasma, and Neorickettsia.
UR - http://www.scopus.com/inward/record.url?scp=0346848864&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0346848864&partnerID=8YFLogxK
U2 - 10.1128/JCM.41.12.5434-5441.2003
DO - 10.1128/JCM.41.12.5434-5441.2003
M3 - Article
C2 - 14662922
AN - SCOPUS:0346848864
SN - 0095-1137
VL - 41
SP - 5434
EP - 5441
JO - Journal of clinical microbiology
JF - Journal of clinical microbiology
IS - 12
ER -