为筛选柔嫩艾美耳球虫(Eimeria tenella)候选疫苗分子,注释并克隆E.tenella人侵相关蛋白EtMAl和EtMA2的编码基因,并对其序列结构进行分析。本研究利用生物信息学和比较基因组学技术注释获得EtMAl和EtMA2的编码基因序列,以E.tenella(广东株)裂殖子总RNA为模板,利用RT—PCR的方法扩增获得EtMAl和EtMA2的开放阅读框(ORF)序列。利用在线生物信息学软件分析EtMAl和EtMA2的序列结构及与顶复门其他虫属的进化关系。结果显示,EtMAl和EtMA2ORF全长分别为1617和1725bp,分别编码全长为539个氨基酸和575个氨基酸的多肽片段。二者与TgAMAl相似性分别为30%和26%。EtMAl和EtMA2同属于E.tenella顶膜抗原超家族蛋白,可能在虫体对宿主的入侵过程中起着关键作用,这也为防控鸡球虫病提供了新的候选疫苗分子。
To screen the novel candidate vaccine for E. tenella, the genes of EtMA1 and EtMA2 associated with parasites invasion were cloned and the sequences were analyzed. The sequence of EtAMA1 cloned in our laboratory were analyzed by the bioinformatics tool and the technology of comparative genomics, and the gene sequences encoding EtMA1 and EtMA2 were acquared. The EtMA1 and EtMA20RF were amplified by RT-PCR from E. tenella (Guangdong strain) mero- zoites total RNA template. The sequence structures of EtMA1 and EtMA2, and the evolution re- lationship with other apicomplexan parasites were analyzed by bioinformatics software on line. The result showed that the ORF of EtMA1 and EtMA2 were 1 617 and 1 725 bp in full length, and coding the peptides of 549 and 575 aa in length. The amino acid sequences were highly con- served with E. tenella apical membrane antigen (EtAMA1) and TgAMA1. Both of the EtMA1 and EtMA2 were superfamily proteins of AMA, and which maybe play a key role in parasites in vasion of host cells, and both of which will be as novel candidate vaccine for E lenella.