论文部分内容阅读
目的为了进一步完善恙虫病立克次体的株型鉴定技术，并且为临床提供诊断试剂。方法本文采用ＮＰＣＲ技术，参考并自行设计两对寡核苷酸引物（Ｐ１、Ｐ２、Ｐ３、Ｐ４），从恙虫病立克次体Ｋａｒｐ株基因组ＤＮＡ中特异扩增出编码Ｓｔａ５６抗原的部分基因片段，经纯化后用ＢａｍＨⅠ和ＨｉｎｄⅢ双酶切后，克隆到质粒ＰＵＣ１８中，转化大肠杆菌ＴＧ１，经ＰＣＲ和双酶切鉴定。结果与结论构建了重组质粒ＰＵＣ１８－ＲＫ，从而为该基因片段的表达奠定了基础。
Objective In order to further improve the Rickettsia tsutsugamushi plant type identification technology, and provide clinical diagnostic reagents. Methods In this paper, two pairs of oligonucleotide primers (P1, P2, P3, P4) were designed and synthesized by NPCR technique. The partial gene fragment encoding Sta56 antigen was amplified from genomic DNA of Karp strain of Rickettsia tsutsugamushi , After purification, digested with BamHⅠand HindⅢ, cloned into plasmid pUC18 and transformed into E. coli TG1, identified by PCR and double enzyme digestion. RESULTS AND CONCLUSION: The recombinant plasmid PUC18-RK was constructed, which laid the foundation for the expression of this gene fragment.