鸡堆型艾美耳球虫新基因的克隆与生物信息学分析

中国畜牧兽医

鸡堆型艾美耳球虫新基因的克隆与生物信息学分析

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

(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)

修回日期:2014-04-18 出版日期:2014-07-20 发布日期:2014-08-21
通讯作者: 黄兵(1958—），男，四川人，学士，研究员，研究方向：动物寄生虫学与寄生虫防治。E-mail:huangbing232@163.com
作者简介:阎晓菲(1983—），女，山东人，硕士，讲师，研究方向：球虫生物学。
基金资助:
国家科技基础条件平台建设项目(2005DKA21205-4)。

Cloning and Bioinformatics Analysis of cDNA Expression Libraries in Eimeria acervulina

YAN Xiao-fei¹, HUANG Bing², HAN Hong-yu², YANG Bin¹, YUE Cheng³, ZHAO Qi-ping², JIANG Lian-lian², DONG Hui²

(1.Science and Technology Department, Xinjiang Agricultural University, Urumqi 830052, China；2. Key Laboratory for Animal Parasitology of Ministry of Agriculture, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China；3. College of Animal Medicine, Xinjiang Agricultural University, Urumqi 830052, China)

Revised:2014-04-18 Online:2014-07-20 Published:2014-08-21

摘要/Abstract

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。

关键词: 鸡; 球虫; 堆型艾美耳球虫; 新基因; 克隆; 生物信息学

Abstract: Analysing the new gene of E. acervulina sporulated oocysts stage by clone and bioinformatics technology. We acquired 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 showed the EST sequence was a new gene of E. acervulina sporulated oocysts stage, the GenBank ID was EU590120. The EST sequence 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 + Glu), 8 basic amino acids(Arg and Lys)，molecular formula was C₇₆₁H₁₂₂₃N₁₉₉O₂₁₉S₁₂, the total average hydrophilic(GRAVY) was 0.731. The signal peptide analysis showed that the ORF encoding protein was an secretory protein containing a signal peptide and four transmembrane structures, its the amino acids residues between 105 to 115, 28 to 32 and 132 to 138 had strong hydrophobicity. At the same time, the protein contains one protein kinase C phosphorylation sites, one N-glycosylation sites, two casein kinase Ⅱ phosphorylation site, four N-end nutmeg acetoxylation site. The bioinformatics analysis showed that the ORF encoding protein possess several biological function sites and potential antigen epitopes.

Key words:

chicken；coccidia；Eimeria acervulinamso-bidi-font-family: 宋体">；new gene；cloning；bioinformatics

阎晓菲，黄兵，韩红玉，杨斌，岳城，赵其平，姜连连，董辉. 鸡堆型艾美耳球虫新基因的克隆与生物信息学分析[J]. 中国畜牧兽医.

YAN Xiao-fei, HUANG Bing, HAN Hong-yu, YANG Bin, YUE Cheng, ZHAO Qi-ping, JIANG Lian-lian, DONG Hui. Cloning and Bioinformatics Analysis of cDNA Expression Libraries in Eimeria acervulina[J]. China Animal Husbandry & Veterinary Medicine.

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