堆型艾美球虫广东株3-1E基因的克隆与序列分析

Cloning and sequence analysis of 3-1E gene of Eimeria acervulina isolated from Guangdong Province

根据GenBank中登录的堆型艾美球虫31E基因序列,设计了3条引物,以广东株堆型艾美球虫裂殖子总RNA为模板,利用反转录聚合酶链反应(RTPCR)扩增获得了31E基因部分片段,将这一片段克隆至pGEMTEasy载体中,经PCR、限制性内切酶分析和克隆片段的序列测定、比较,证实了克隆片段的可靠性。序列比较发现,所克隆的基因片段与Eimeriaacervulina美国株(US)、E.acervulinaQH株31EcDNA的核苷酸同源性分别为99.0%和99.2%,推导氨基酸的同源性分别为98.2%和97.6%。

Using the total RNA of merozoites of Eimeria acervulina isolated from Guangdong Pro- vince as template, a partial segment of 3-1E gene was amplified by RT-PCR. The gene fragment 682bp in length was cloned into pGEM-T Easy vector, and the recombinant plasmid was identified by PCR, restriction enzyme analysis and sequencing. The homology analysis revealed that the nucleotide homology between the 3-1E gene of the E. acervulina Guangdong strain and that of the US strain and QH strain was 99.0% and 99.2% respectively, whereas the deduced amino acid homology was 98.2% and 97.6% respectively.