斯氏艾美耳球虫ADF基因部分序列的克隆与分析

Cloning and sequence analysis of partial sequences of ADF genes from Eimeria stiedai

为了获得我国斯氏艾美耳球虫长春株ADF基因并分析其与柔嫩艾美耳球虫长春株相应序列的同源性,试验根据GenBank中公布的柔嫩艾美耳球虫ADF基因序列设计引物,采用RT-PCR方法获得我国斯氏艾美耳球虫长春株ADF基因部分序列,然后将其克隆到pMD18-T载体中,应用DNAStar软件对测得序列进行序列分析。结果表明:斯氏艾美耳球虫长春株ADF基因大小为357 bp;经DNAStar分析,我国斯氏艾美耳球虫长春株与柔嫩艾美耳球虫长春株ADF核苷酸序列和氨基酸序列同源性均为99.2%。说明ADF基因在进化过程中高度保守。

To obtain the ADF genes of Changchun strains of E. stiedai and analyze its homology of the corresponding sequences of Changchun strains of E. tenella, a pair of primers was designed according to the ADF sequences of E. tenella published in the GenBank. The partial se quences of ADF genes from Changchun strains of E. stiedai were obtained by RT PCR. The PCR product was subcloned to the pMD18 T vector, and the measured sequences were used to be sequence analysis by DNAStar software. The results showed that the ADF genes from Chang ehun strains of E. stiedai were 357 bp in length. The DNAStar analysis showed that the homologies of ADF nucleotide and amino acid sequences from Changchun strains between E. stiedai and E. tenella were all 99.2%. It indicates that ADF genes are highly conservative in the evolution process.