鸡堆型艾美耳球虫新基因的克隆与生物信息学分析  

Cloning and Bioinformatics Analysis of cDNA Expression Libraries in Eimeriaacervulina

作　　者：阎晓菲[1] 黄兵[2] 韩红玉[2] 杨斌[1] 岳城[3] 赵其平[2] 姜连连[2] 董辉[2] 

机构地区：[1]新疆农业大学科学技术学院,新疆乌鲁木齐830052 [2]中国农业科学院上海兽医研究所,农业部动物寄生虫学重点实验室,上海200241 [3]新疆农业大学动物医学学院,新疆乌鲁木齐830052

出　　处：《中国畜牧兽医》2014年第7期49-53,共5页China Animal Husbandry & Veterinary Medicine

基　　金：国家科技基础条件平台建设项目(2005DKA21205-4)

摘　　要：试验旨在对堆型艾美耳球虫（Eimeria acervulina）孢子化卵囊新基因EST序列进行克隆与生物信息学分析。通过RACE技术扩增从E．acervulina cDNA表达文库中筛选获得的EST序列，得到EST全长cDNA序列，利用多种生物信息学分析软件对其编码蛋白进行分析和预测。结果表明，该EST序列为E．acervulina孢子化卵囊阶段新基因，GenBank登录号为EU590120；该基因含1个492 bp的开放阅读框，编码163个氨基酸，理论分子质量为17．049 ku，等电点为6．69，酸性氨基酸8个，碱性氨基酸8个，分子式为C761H1223N199O219S12总平均亲水性（GRAVY）为0．731；该蛋白有信号肽，为分泌性蛋白；具有4个跨膜结构，在105～115、28～32和132～138位之间有很强的疏水性；具有1个蛋白激酶C磷酸化位点、1个N-糖基化位点、2个酪蛋白激酶Ⅱ磷酸化位点、4个N端豆蔻酰基化位点。因此根据生物信息学全面预测和分析，发现该基因所编码的蛋白具有较多的生物学功能位点和潜在的抗原表位区域。Analysing the new gene of E. acervulina sporulated oocysts stage by clone and bioinformatics technology. Weacquired the full-length cDNA of EST sequence by RACE technique based on the cDNA expression library of E. acervulina.The cDNA of EST sequence coded protein was analysised and predicted by several bioinformatics software, the result showedthe EST sequence was a new gene of E. acervulina sporulated oocysts stage, the GenBank ID was EU590120. The EST se-quence including a 492 bp open reading frame which coded 163 amino acids residues, the theory of molecular weight was 17. 049 ku,isoelectric point was 6.69, 8 acidic amino acids （Asp q- Glu）, 8 basic amino acids （Arg and Lys）, molecular formula wasC7nl Hm3 N1990219 Slz, the total average hydrophilic （GRAVY） was 0. 731. The signal peptide analysis showed that the ORFencoding protein was an secretory protein containing a signal peptide and four transmembrane structures, its the amino acidsresidues between 105 to 115, 28 to 32 and 132 to 138 had strong hydrophobicity. At the same time, the protein contains oneprotein kinase C phosphorylation sites, one N-glycosylation sites, two casein kinase Ⅱphosphorylation site, four N-end nut-meg acetoxylation site. The bioinformatics analysis showed that the ORF encoding protein possess several biological functionsites and potential antigen epitopes.

关 键 词：鸡 球虫 堆型艾美耳球虫 新基因 克隆 生物信息学 

分 类 号：S852.723[农业科学—基础兽医学]