柔嫩艾美耳球虫河北株SOSO7基因克隆与序列分析  

Cloning and Sequence Analysis of SO7 Gene of Eimeria tenella Hebei Strain

作　　者：闫艳娟 庞洪泽 李蕴玉[1] 张东林[1] 李佩国[1] 张香斋[1] 张召兴[1] 唐梦虎 YAN Yanjuan;PANG Hongze;LI Yunyu;ZHANG Donglin;LI Peiguo;ZHANG Xiangzhai;ZHANG Zhaoxing;TANG Menghu(Hebei Province Key Laboratory of Veterinary Preventive Medicine,Hebei Normal University of Science ＆ Technology,Qinhuangdao,Hebei 06660)

机构地区：[1]河北科技师范学院河北省预防兽医学重点实验室,河北秦皇岛066600

出　　处：《中国家禽》2018年第17期22-25,共4页China Poultry

基　　金：河北省现代农业产业技术体系蛋肉鸡产业创新团队(HBCT2018150206);2017年河北省硕士研究生创新资助项目(CXZZSS2017158)

摘　　要：为分析柔嫩艾美耳球虫(Eimeria tenella)河北株SO7基因序列遗传变异性,试验根据GenBank发表的SO7序列设计引物,利用RT-PCR技术克隆出柔嫩艾美耳球虫河北株SO7基因,将SO7基因克隆入pMD18-T载体中,进行测序、序列分析及表达蛋白结构的预测。结果显示:柔嫩艾美耳球虫河北株SO7基因编码215个氨基酸,具有一个开放阅读框,大小为645 bp;与GenBank中发表的美国分离株(LS18)、扬州分离株(YZ)、海南分离株(HN)、广东分离株(GD)、北京分离株(BJ)SO7核苷酸序列的同源性分别为99.4%、99.5%、99.7%、99.5%、99.4%,所推导的氨基酸序列的同源性分别为98.6%、99.1%、99.1%、98.6%、98.1%。从进化关系来看,同种间SO7基因相比,柔嫩艾美耳球虫河北株与HN株同属一个分支,遗传关系较近;与GD株、BJ株的遗传关系相对较远。蛋白结构预测显示蛋白的相对分子质量为52.20 ku,理论等电点(PI)为5.08,不稳定系数为52.73,为不稳定蛋白;在1~20个氨基酸具有信号肽;有5个疏水位点;没有跨膜结构。综上所述,柔嫩艾美耳球虫河北株SO7基因序列遗传变异性不明显,可以作为构建核酸疫苗候选基因。In order to analyze the genetic variability of SO7 gene sequence of Eimeria tenella Hebei （HB）strain, a pair of primers were designed which was based on the published ORF of E. tenella SO7 gene. The gene was amplified by RT-PCR from HB strain, then cloned into pMD18-T vector for sequence analysis and structure prediction of the expressed protein. The results showed that the gene was 645 bp, including an open reading frame and encoding 215 amino acids. The nucleotide sequence homology of SO7 gene of HB strain was respectively 99.4%, 99.5%, 99.7%, 99.5% and 99.4% with the American isolate （LS18）, Yangzhou isolate （YZ）, Hainan isolate （HN）, Guangdong isolate （GD） and Beijing isolate （B J）published in GenBank and the amino acid sequence homology with the genes encoding wasrespectively 98.6%, 99.1%, 99.1%, 98.6% and 98.1%. From the evolutionary point of view, the HB strain and HN strain belonged to the same branch and had genetic relationships with each other, and the genetic relationship with GD strain and BJ strain was relatively distant. The prediction of protein structure showed that the molecular weight of encoded protein is 52.20 ku, the theoretical isoelectric point was 5.08. The protein was an unstable protein and unstable coefficient was 52.73. The signal peptide was located in 1 to 20 amino acids. There were 5 hydrophobic sites, no transmembrane structure. In summary, the genetic variability of SO7 gene sequence of the E. tenella HB strain was not obvious, and it could be used as a gene candidate for constructing a nucleic acid vaccine.

关 键 词：柔嫩艾美耳球虫河北株 SO7基因 序列分析 蛋白结构预测 

分 类 号：S855.9[农业科学—临床兽医学]