毒害艾美耳球虫配子体gam22基因的克隆与序列分析

Cloning and sequence analysis of gam22 gene from Eimeria necatrix

提取毒害艾美耳球虫(En)配子体总RNA,应用RT-PCR进行gam22基因的克隆,测序后对其进行序列分析。结果表明,Engam22基因全长为713 bp,其中包含一个完整的开放阅读框,大小为561 bp,编码186个氨基酸,含有一个富含组氨酸和脯氨酸的特征性结构域;与柔嫩艾美耳球虫(Et)配子体Etgam22基因序列进行比对,同源性为97.7%。此配子体蛋白基因具有较高的保守性,可以作为抗球虫疫苗的候选基因。

Total RNA isolated from Eimeria necatrix gametocytes was utilized as template for RT-PCR amplification of gam22 gene, which encodes a gametocyte protein. Then the gene was sequenced. The results showed that the gain22 gene was 713 bp in length, which contained a complete open reading frame of 561 bp, encoding 186 amino acids. Its encoded protein contains a characteristic domain rich in histidine and proline. The gene had 97.7% identity to that of gam22 of E. tenella, which presented a potential target for future recombinant subunit vaccines against coccidiosis.