长角血蜱卵cDNA表达文库的构建.docVIP

长角血蜱卵cDNA表达文库的构建 刘光远，田占成，才学鹏，李知新，谢俊仁，王 路，张 林，龚真莉 （中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室甘肃省动物寄生虫病重点实验室·ml-1。获得了目的基因，其序列与GenBank中的长角血蜱促卵泡激素（DQ248886）的核苷酸序列的同源性为97.8%。【结论】成功构建了长角血蜱卵cDNA表达文库。 关键词：长角血蜱；卵；cDNA表达文库；构建 Construction of cDNA Expression Library from Eggs of Haemaphysalis longicornis LIU Guang-yuan, TIAN Zhan-cheng, CAI Xue-peng, LI Zhi-xin, XIE Jun-ren, WANG Lu, ZHANG Lin, GONG Zhen-li (Key Laboratory of Veterinary Parasitology of Gansu Province/State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046) Abstract: 【Objective】 cDNA expression library from eggs of Haemaphysalis longicornis for further screening and cloning of the potential candidate antigenic genes. 【Method】 Total RNA was isolated from eggs of H. longicornis mRNA were purified, and cDNAs were synthesized and ligated to λSCREEN vector. The recombinant phage DNA was packaged by using Phagemark packaging extracts and then transfected to E. coli ER1647 to obtain the cDNA expression library. 【Result】The size of the primary cDNA library was 1.38×106 PFU with titer of the amplified cDNA library of 2×109 PFU·ml-1. A full length cDNA encoding follistatin-related protein was cloned from the cDNA library by PCR. 【Conclusion】The cDNA expression library from eggs of H. longicornis was successfully constructed. Key words: Haemaphysalis longicornisEggs; cDNA expression library; Construction 0 引言 【研究意义】蜱在摄食过程中，不仅给宿主造成直接损害，而且传播多种病原体由蜱传播的新老疾病在对畜牧业生产造成巨大经济损失的同时，也引发了严重的公共卫生问题。因而，对蜱的控制非常必要[1]。传统的控蜱方法主要是使用杀虫剂，但由于某些杀虫剂会产生抗药性，对畜产品和环境造成污染等，因而抗蜱疫苗的研制显示出较好的前景[2]。为寻找高效、广谱的抗蜱疫苗候选分子、描述蜱与宿主的免疫生物学特性，必须对蜱吸血、病原传播及蜱与宿主之间的相互作用有更加深入的了解[3]。长角血蜱Haemaphysalis longicornis）主要分布在国东北、华北、西北等广大地区，是流行最广泛和最重要的蜱种之一[4]，传播牛、羊泰勒属Theileria）和巴贝斯属Babesia）的多种病原，具有重要的流行病学意义。【前人研究进展】在过去的年，研究蜱与宿主相互作用的免疫生物学技术取得了很大进步。根据宿主对蜱叮咬获得先天性或特异性免疫反应的机制，集中研究了相当多的单个蜱对宿主免疫系统的影响[56]。利用功能基因组和蛋白组学技术加速了对蜱、宿主、病原相互作用的研究步伐[7]。目前通过多种手段已经发现许多蜱的免疫相