Dynamics of Rickettsia-tick interactions:: identification and characterization of differentially expressed mRNAs in uninfected and infected Dermacentor variabilis
Dynamics of Rickettsia-tick interactions:: identification and characterization of differentially expressed mRNAs in uninfected and infected Dermacentor variabilis
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DOI：
10.1046/j.1365-2583.2003.00400.x
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发表时间：
2003-04-01
影响因子：
2.6
通讯作者：
Azad, AF
中科院分区：
文献类型：
--
作者：
Mulenga, A;Macaluso, KR;Azad, AF
为了开始探索立克次体蜱相互作用的分子动力学，使用消减杂交来筛选感染和未感染蒙大拿立克次体的可变革蜱中的mRNA。我们分离了30个cDNA片段，其中22个上调，8个下调立克次体感染。根据11个与已知蛋白质家族相似的cDNA片段的推定同一性，将蜱类遗传因子分为3类，包括蜱类免疫应答因子（α-2巨球蛋白和IgE依赖性组胺释放因子），受体/粘附分子（信号转导和转录激活因子-1/3蛋白抑制剂，网格蛋白衔接蛋白和四跨膜蛋白）和应激反应蛋白（醛糖还原酶、谷胱甘肽-S转移酶、铁蛋白、核小体装配蛋白和细胞周期蛋白A蛋白）。半定量RT-PCR扩增产物的密度分析表明，差异表达的23个遗传因子中的18个，显然代表了78%的一致性，通过消减杂交获得的结果。此外，从蜱血细胞/循环细胞中扩增了23种检测的遗传因子中的17种的mRNA转录本，表明其表达不限于卵巢，并表明可能参与免疫应答。
To begin to explore the molecular dynamics of rickettsial tick interaction, subtractive hybridization was used to screen mRNAs in Rickettsia montanensis -infected and uninfected Dermacentor variabilis . We isolated 30 cDNA fragments, 22 of which were up-regulated and eight were down-regulated in response to rickettsial infection. Based on a putative identity of 11 cDNA fragments with similarity to known protein families, the tick genetic factors have been assigned into three groups including, the tick immune response factors (alpha-2 macroglobulin and IgE-dependent histamine release factor), the receptor/adhesion molecules (the signal transducer and activator of transcription-1/3 protein inhibitor, the clathrin adaptor protein and tetraspanin) and the stress response proteins (aldose reductase, glutathione-S transferase, ferritin, nucleosome assembly protein and cyclin A protein). Density analyses of semiquantitative RT-PCR amplified products demonstrated differential expression for 18 of the 23 tested genetic factors, apparently representing a 78% agreement with results obtained by subtractive hybridization. Additionally, mRNA transcripts of 17 of the 23 tested genetic factors were amplified from tick haemocytes/circulatory cells demonstrating that their expression is not restricted to the ovaries and suggesting a potential involvement in the immune response.