堆型艾美耳球虫保定株裂殖子3-1E基因的克隆与序列分析

Cloning and sequaence analysis of 3-1E gene of merozoites of Eimeria acervulina Baoding strain isolated from Hebei province of China

依据GenBank中收录的堆型艾美耳球虫(Eimeria acervulina,E.acervulina)美国株(US)3-1E基因序列,设计特异性引物,以堆型艾美耳球虫保定株裂殖子基因组RNA为模板,利用反转录-聚合酶链式反应(RT-PCR)扩增获得3-1E基因序列部分片段,将此片段克隆至pGM-T Easy载体中,经PCR、限制性内切酶鉴定和克隆片段的序列测定、比较,结果表明该克隆片段扩增准确、可靠。序列比较发现,此片段与E.acervulina美国株(US)株、E.acervulina QH株cDNA的核苷酸同源性分别为99.4%和99.6%。

Based on the sequence alignment of the 3-1E gene of Eimeria acervulina （E. acervulina）US strain, two pairs of primers were designed and synthesized. Using the total RNA of merozoites of E. acervulina Baoding strain isolated from Hebei province of China as template,a partial segment of 3-1E gene was amplified by RT-PCR. The gene fragment 689 bp in length was cloned into pGM-T Easy vector,and the recombinant plasmid was identified by PCR.restriction enzyme analysis and sequencing. The homology analysis revealed that the nucleotide sequences of the 3-1E gene of the E. acervulina Baoding strain similar to the US strain and QH strain were 99.4% and 99.6% respectively.