检测沃尔巴克氏体编码的 DNA 结合蛋白 HU beta 在蚊子生殖腺中的存在。
Detection of the Wolbachia-encoded DNA binding protein, HU beta, in mosquito gonads.
机构信息
Department of Entomology, University of Minnesota, St. Paul, MN 55108, USA.
出版信息
Insect Biochem Mol Biol. 2013 Mar;43(3):272-9. doi: 10.1016/j.ibmb.2012.12.007. Epub 2012 Dec 31.
Abstract
Wolbachia are obligate intracellular bacteria that cause cytoplasmic incompatibility in mosquitoes. In an incompatible cross, eggs of uninfected females fail to hatch when fertilized by sperm from infected males. We used polyacrylamide gel electrophoresis and tandem mass spectrometry to identify Wolbachia proteins in infected mosquito gonads. These included surface proteins with masses of 25 and 18 kDa and the DNA binding protein, HU beta. Using reverse transcriptase polymerase chain reaction, we showed that the HU gene is transcribed in Wolbachia-infected Culex pipiens and Aedes albopictus mosquitoes. We sequenced HU genes from four Wolbachia strains and compared deduced protein sequences with additional homologs from the databases. Among the Rickettsiales, Wolbachia HU has distinct N- and C-terminal basic/acidic amino acid motifs as well as a pair of conserved, cysteine residues.
摘要
沃尔巴克氏体是一种专性细胞内细菌，会导致蚊子的细胞质不亲和。在不相容的杂交中，未感染的雌性的卵子在被感染雄性的精子受精后无法孵化。我们使用聚丙烯酰胺凝胶电泳和串联质谱法来鉴定感染蚊子生殖腺中的沃尔巴克氏体蛋白。这些蛋白包括分子量为 25 和 18 kDa 的表面蛋白和 DNA 结合蛋白 HUβ。通过逆转录聚合酶链反应，我们表明 HU 基因在感染沃尔巴克氏体的库蚊和白纹伊蚊中转录。我们从四种沃尔巴克氏体菌株中测序了 HU 基因，并将推导的蛋白质序列与数据库中的其他同源物进行了比较。在立克次氏体目中，沃尔巴克氏体 HU 具有独特的 N 端和 C 端碱性/酸性氨基酸基序以及一对保守的半胱氨酸残基。
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