摘要
目的构建弓形虫pcDNA3-ROP2-p30-HSP70真核表达重组质粒。方法根据HSP70基因序列,设计合成HSP70基因的引物,PCR扩增HSP70基因片段,克隆入pMD18-T载体,再经酶切、连接等,亚克隆至pcDNA3-ROP2-p30表达重组质粒中,并进行酶切、PCR和测序鉴定。结果 PCR扩增出长度为916bp的HSP70基因片段,将其克隆到pMD18-T中,成功亚克隆获得pcDNA3-ROP2-p30-HSP70表达重组质粒。测序结果显示,pcDNA3-ROP2-p30-HSP70表达重组质粒包含了HSP70蛋白基因完整序列。结论成功构建弓形虫pcDNA3-ROP2-p30-HSP70真核表达重组质粒,为下一步核酸疫苗的研究奠定了基础。
Objective To construct a recombinant eukaryotic expression plasmid pcDNA3-ROP2-p30-HSP70 of Toxoplasma gondii.Methods The primers of HSP70 gene were designed and the HSP70 gene were amplified by PCR,then the HSP70 gene was inserted into cloning vector pMD18-T.Then the inserted HSP70 gene fragment was subcloned to pcDNA3-ROP2-p30 recombinant by digesting with restrictive enzymes and connection reactions.The positive clone was identified by restrictive enzyme digestion and PCR amplification.Results A total of 916 HSP70 fragment genes were amplified and inserted into the eukaryotic expression vector pcDNA3-ROP2-p30.The cloning and eukaryotic expression recombinant plasmid was correctly constructed.The results showed that HSP70 protein in the recombinant plasmid contained the complete gene sequence.Conclusions The recombinant plasmid pcDNA3-ROP2-p30-HSP70 is successfully constructed.This will pave the way for further study of DNA vaccination of T.gondii.
出处
《中国血吸虫病防治杂志》
CAS
CSCD
北大核心
2010年第4期329-332,共4页
Chinese Journal of Schistosomiasis Control
基金
山东省医学科学院科研基金(2006-08)
