柔嫩艾美耳球虫ZJ株5401基因的克隆和序列分析

Clone and Sequence Analysis of Gene5401 of Eimeria Tenella ZJ Strain

利用RT PCR方法扩增出了柔嫩艾美耳球虫(E.tenella)ZJ(浙江)株的子孢子表面抗原5401基因。把这一基因片段克隆到PGEM T克隆载体上,得到的阳性克隆经PCR鉴定及酶切分析,结果表明重组子(pGEM T 5401)中含有5401基因,该序列全长为864bp,序列本身是一个开放阅读框,将克隆得到的基因与国外报道的5401基因比较,有两个碱基发生有义突变,引起两个氨基酸发生突变,核苷酸同源性为99 8%,氨基酸同源性为99 3%。克隆出的5401基因可以用于将来重组疫苗的研究。

A sporozoite surface antigen 5401 gene of E.tenella was cloned by RTPCR from the synthesized first strand cDNA.The PCR products were analysed by agarose electrophoresis and then cloned into pGEMT vector successfully.The specific recombinant plasmid was identified with restriction endonuclease analysis and sequenced.The result indicated the 5401 gene was 864bp in length and the sequence itself is an open reading frame which encodes a peptide of 287 amino acid residues.Comparison of the 5401 gene we gained with the previously reported 5401 gene indicated two genes had a high homology,and the 5401 gene we gained changed 2 nucleotides compared with the reported 5401 gene,which indicited 2 amino acid changed.