细粒棘球蚴内蒙株疫苗候选基因EG95克隆

Cloning and sequence analyzing of vaccine candidated gene EG95 from Echinococcus granulosus

目的克隆细粒棘球蚴内蒙株EG95基因,并进行序列分析。方法根据GenBank中细粒棘球蚴EG95-1基因DNA序列设计引物,提取细粒棘球蚴基因组DNA,PCR扩增目的基因,目的基因克隆到pMD19-T载体,经PCR、酶切及测序鉴定后,进行序列分析。结果克隆到的EG95-NM基因序列长1262bp,与EG95-1基因DNA序列同源性为98%。结论克隆到细粒棘球蚴内蒙株EG95基因,序列分析表明,EG95-NM基因属于EG95基因家族并具有地理株特点。

Objective To clone the EG95 gene of Echinococcus granulosus in Inner Mongolia and analyze the gene nucleotide sequence.Methods According to the sequence of EG95-1 gene from genbank,a pair of specific primers were designed.The genomic DNA was extracted from Echinococcus granulosus.EG95-NM gene was amplified with the primers using PCR and cloned into pMD19-T plasmid.The recombinant plasmid was characterized with PCR,enzyme digestion and sequencing.The EG95-NM nucleotide sequence was analyzed by DNAstar biosoftware.Results The length of the EG95-NM nucleotide sequence was 1262bp.The EG95-NM gene sequence shared 98% identity with EG95-1 gene.Conclusion The sequence analyzing reveals that the EG95-NM gene belongs to EG95 gene family.