两种兔艾美耳球虫顶质体rpoB和TufA基因的扩增及序列分析

为了探讨顶质体序列在顶复门寄生虫遗传进化研究中的意义,应用PCR技术扩增了兔维氏艾美耳球虫(Eimeria vejdovskyi)和无残艾美耳球虫(E.irresidua)顶质体rpoB和TufA基因序列,并利用生物学软件与GenBank中收录的顶复门寄生虫相关序列进行比对和进化分析。结果表明:扩增的两种兔艾美耳球虫rpoB和TufA基因片段的大小均相同,分别为469 bp和481 bp,两种球虫rpoB和TufA基因种间的差异分别为6.8%和0.2%,rpoB基因差异相对较大,而TufA基因差异较小。因此,rpoB基因序列可以作为兔艾美耳球虫种间的遗传标记,用得到的rpoB基因序列为参考与其他顶复门寄生虫进行进化分析进一步证明了该结论,实验首次报道兔艾美耳球虫顶质体rpoB和TufA基因的部分序列,为兔艾美耳球虫的鉴定和进化分析提供参考依据,也为顶复门顶质体的深入研究奠定了基础。

PCR Amplification and Sequence Analysis of Apicoplast rpoB and TufA Genes of Two Eimeria spp. in Rabbits

The objective of the present study was to examine sequence variation in apicoplast rpoB and TufA genes of Eimeria vejdovskyi and E.irresidua of rabbits.The partial sequences of rpoB and TufA genes were amplified from the two Eimeria spp. by polymerase chain reaction （PCR）,and then sequenced directly.The obtained sequences were compared with published relevant sequences of Apicomplexan parasites in GenBank to analyze sequence differences. The results showed that the length of the partial rpoB sequence was 469 bp,and the partial Tufa sequence was 481 bp.Sequence differences in rpoB gene and TufA gene were 6.8% and 0.2% between the two rabbits Eimeria species respectively.RpoB sequence provided novel genetic markers for the identification of Eimeria species, which was supported by phylogenetic analysis of apicoplast rpoB sequences.The present study was the first to report partial apicoplast rpoB and TufA gene sequences of rabbit Eimeria spp. ,which provided useful genetic markers for species identification and phylogenetic analysis of rabbit Eimeria.