The molecular and biological analysis of ixodid ticks histamine release factors
The molecular and biological analysis of ixodid ticks histamine release factors
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DOI：
10.1007/s10493-005-3261-8
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发表时间：
2005-12-01
影响因子：
2.2
通讯作者：
Azad, AF
中科院分区：
文献类型：
--
作者：
Mulenga, A;Azad, AF
我们先前描述了一种Dermacentor variibialis（DV）cDNA，其编码普遍表达的和蜱唾液分泌的功能性组胺释放因子（HRF）同源物。本研究利用DVHRF开放阅读框序列设计的基因特异性引物，从木蜱、D. andersoni（DA）、黑腿蜱、南方牛蜱、微小牛蜱（BM）和孤星星蜱Amblyomma americanum（AA）。在核苷酸水平上，DAHRF、AAHRF和BMHRF与DVHRF的同源性分别为98.89%和84%，而预测的多肽序列的同源性分别为98.96%和91%。系统发育，蜱HRF分支，而不同的（100%自举值），是密切相关的其他节肢动物，但远相关的脊椎动物和原生动物的分支。与序列相似性分析一致，DVHRF特异性北方印迹探针在所有RNA印迹上杂交了相似的900碱基对（bp）mRNA条带。小鼠抗E.大肠杆菌表达的重组（r）DVHRF、交叉反应的杆状病毒表达的非融合rAAHRF、rDAHRF和rBMHRF。幼虫和若虫RNA的北方印迹分析显示，DVHRF mRNA在未成熟和成熟蜱中均表达，表明其转录不受发育调节。与其他生物体的rHRF/TCTP蛋白不同，钙结合功能可能不保守蜱HRF同源物所揭示的（CaCl 2）-Ca-45 +覆盖试验。DVHRF及其直系同源物的表观全球表达使该蛋白质家族成为开发针对多个蜱物种的新型蜱控制策略的理想靶抗原。
We previously described a Dermacentor varibialis (DV) cDNA that encodes a ubiquitously expressed and tick saliva-secreted functional histamine release factor (HRF) homolog. In this study gene specific primers based on DVHRF open reading frame nucleotide sequence were utilized to amplify three orthologs, from the wood tick, D. andersoni (DA), the black legged tick, the southern cattle tick, Boophilus microplus (BM) and the lone star tick, Amblyomma americanum (AA). At nucleotide level, sequence comparisons revealed 98 89 and 84% similarity to DVHRF for DAHRF, AAHRF and BMHRF, respectively, while predicted polypeptide comparisons revealed 98, 96 and 91% similarity for DAHRF, AAHRF and BMHRF respectively. Phylogenetically, the tick HRF clade, while distinct (100% bootstrap value), is closely related to other arthropods, but distantly related to vertebrate and protozoan clades. Consistent with sequence similarity analysis, a DVHRF-specific northern blotting probe hybridized a similar to 900 base pair (bp) mRNA band on all RNA blots. Likewise a mouse polyclonal antibody to E. coli-expressed recombinant (r) DVHRF, cross-reacted baculovirus-expressed non-fusion rAAHRF, rDAHRF, and rBMHRF. As revealed by northern blotting analysis of larvae and nymph RNA, DVHRF mRNA is expressed in both immature and mature ticks indicating that its transcription is not developmentally regulated. Unlike rHRF/TCTP proteins of other organisms, the calcium-binding function may not be conserved for tick HRF homologs as revealed by the (CaCl2)-Ca-45 + overlay assay. Apparent global expression of DVHRF and its orthologs make this protein family an ideal target antigen for development of novel tick control strategies targeting multiple tick species.