摘要
目的 获取犬贾第虫核糖体 16sRNA基因序列。方法 从犬贾第虫包囊中 ,快速提取DNA ,并以DNA为模板 ,利用热启动PCR技术扩增出一 6 11bp的预计大小的基因片段 ,纯化回收后 ,与PMD18 T载体连接转化到DH5a大肠杆菌中 ,筛选到阳性克隆经PCR、EcoRⅠ和HindⅢ双酶切鉴定 ,并用双脱氧链末端终止法测定DNA序列 ,登陆BLAST进行同源性比较和分析。结果 获得了长度为 6 11bp的基因序列 ,并发现与犬贾第虫核糖体的同源性最高达 99%。结论 获得了犬贾第虫核糖体 16sRNA的部分基因序列 ,为研究其在核酸进化领域中的地位和PCR法快速诊断贾第虫病提供了理想的基因材料。
Objective To clone the partial 16sRNA gene of Giardia canis. Method DNA were prepared from the cysts of G.canis.A 611bp gene fragment was amplified from the G.canis genomic DNA with specific primers,and cloned into the PMD18 T vector.Results The 611bp gene fragment was amplified and cloned.There is a homology of 99% comparing with the previously reported gene. Conclusion The partial 16sRNA gene was isolated and cloned from G.canis,which provided ideal gene for studying its importance in the nucleus evolution and diagnosing giardiasis with PCR approach.
出处
《热带医学杂志》
CAS
2002年第4期324-326,共3页
Journal of Tropical Medicine
