A serine proteinase inhibitor (serpin) from ixodid tick Haemaphysalis longicornis; cloning and preliminary assessment of its suitability as a candidate for a tick vaccine.
A serine proteinase inhibitor (serpin) from ixodid tick Haemaphysalis longicornis; cloning and preliminary assessment of its suitability as a candidate for a tick vaccine.
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DOI：
10.1016/s0264-410x(03)00167-1
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发表时间：
2003-06
期刊：
影响因子：
5.5
通讯作者：
M. Sugino;S. Imamura;A. Mulenga;M. Nakajima;A. Tsuda;K. Ohashi;M. Onuma
中科院分区：
文献类型：
--
作者：
M. Sugino;S. Imamura;A. Mulenga;M. Nakajima;A. Tsuda;K. Ohashi;M. Onuma
利用cDNA末端快速扩增法（RACE）和基于丝氨酸蛋白酶抑制剂保守氨基酸基序设计的引物（NAVYFKG）克隆了一个1350 bp的cDNA，该cDNA编码一个378多肽，与已知的几种丝氨酸蛋白酶抑制剂具有高度的序列相似性。我们将该基因命名为长角血蜱serpin-1（HLS 1）。北方印迹和逆转录（RT）-PCR分析来自未进食或部分进食的完整蜱以及解剖的蜱器官的总RNA显示，在缓慢进食阶段（附着后24- 48 h）期间，仅在蜱中肠中，由血粉进食诱导HLS 1 mRNA的转录。用大肠杆菌表达的重组HLS 1（rHLS 1）免疫家兔，其蜱若虫和成蜱的死亡率分别为43.9%和11.2%。兔抗蜱唾液的多克隆抗体不与rHLS 1发生反应，表明在蜱取食过程中，原生HLS 1不会分泌到宿主体内。rHLS 1可能是鸡尾酒抗蜱疫苗的潜在候选者。
Rapid amplification of the cDNA ends (RACE) and primers designed based on a conserved serpin amino acid motif (NAVYFKG) were used to clone a 1350bp cDNA which encodes a 378 polypeptide with high sequence similarity to several known serpins. We have named this gene as Haemaphysalis longicornis serpin-1 (HLS1). Northern blotting and reverse transcription (RT)-PCR analysis of total RNA from unfed or partially fed whole ticks as well as dissected tick organs revealed that transcription of HLS1 mRNA was induced by blood meal feeding during the slow feeding phase (24–48h post-attachment) only in the tick midguts. Vaccination of rabbits with recombinant HLS1 (rHLS1) expressed in Escherichia coli resulted in 43.9 and 11.2% mortality of nymph and adult ticks which were fed on immunized rabbits. Polyclonal rabbit antibodies to tick saliva did not react with rHLS1, suggesting that native HLS1 was not secreted into the host during tick feeding. rHLS1 could be a potential candidate for a cocktail anti-tick vaccine.