长角血蜱半乳糖凝集素的分子特征分析
首发时间:2015-09-11
摘要:半乳糖凝集素通过与大批糖缀合物的结合,介导细胞-细胞之间及细胞-胞外基质之间的相互作用。长角血蜱是家畜巴贝斯虫病的媒介及一些人传染病的储存宿主,研究该蜱与其传播的病原间的相互作用将有助于家畜和人的疾病防治。本实验用PCR法从该蜱cDNA中克隆了一个半乳糖凝集素的同源基因(HlGalec)。该基因编码了一条由323个氨基酸组成的多肽链,预测分子量为36 kDa;结构域分析显示该多肽链具有两个串联的糖识别结构域且含有半乳糖凝集素家族中典型的基序。逆转录PCR检测发现HlGalec的mRNA在所有发育期和多种组织器官中均存在。用毕赤酵母表达的重组HlGalec(rHlGalec)有很强的红血细胞凝集作用及与乳糖亲合的活性。rHlGalec免疫的鼠血清能在免疫印迹试验中识别天然HlGalec蛋白,其表观分子量与理论值相符,免疫组化试验从翻译水平进一步证实HlGalec基因在多种器官中均有表达,在成蜱中肠和血细胞中尤为丰富。rHlGalec与各种不同结构的糖缀合物的亲和谱、HlGalec在蜱和病原相互作用中扮演的角色有待进一步研究。
关键词: 分子特征 长角血蜱 半乳糖凝集素 红血细胞凝集作用 乳糖亲和性
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Molecular characterization of a galectin from Haemaphysalis longicornis
Abstract:Galectins mediate cell-cell and cell-extracellular matrix interaction by binding to a wide array of glycoconjugates. In this paper, a galectin was cloned from the ixoidid tick Haemaphysalis longicornis (HlGalec), a natural vector or reservoir of animal babesiosis and some human infectious diseases by PCR. HlGalec consists of 323 amino acids with a molecular mass of 36 kDa, and possesses two tandem-repeated carbohydrate recognition domains containing typical motif conserved in galectin family. Hlgalec was expressed both transcriptionally and translationally in all development stages and in multiple organs and was abundant in the midgut of the adult tick. A yeast-expressed recombinant HlGalec (rHlGalec) exhibited strong hemagglutination and lactose-binding activities.
Keywords: molecular characterization Haemaphysalis longicornis galectose haemagglutination lactose-binding activity
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