摘要
目的 构建 pcDNA3-HBsAg -GRA1真核表达质粒 ,为提高弓形虫DNA疫苗保护性做出新的探索 ,并期望“一 / +-苗两用”。方法 采用PCR分别扩增HBsAg、GRA1;经BclⅠ、EcoRⅠ双酶切HBsAg ;EcoRⅠ、XhoⅠ双酶切GRA1;BamHⅠ、XhoⅠ双酶切载体 pcDNA3 0 ;HBsAg、GRA1、pcDNA3 0双酶切纯化产物与T4DNA连接酶在 2 2℃连接过夜 ,用双酶切及双脱氧末端终止法测序鉴定重组质粒。结果 成功构建pcDNA3-HBsAg -GRA1真核表达质粒。结论 为进一步研究 pcD NA3-HBsAg -GRA1诱导的保护性免疫奠定了基础。
Objective To construct a eukaryotic expression plasmid encoding hepatitis B surface antigen (HBsAg) and T.gondii GRA1 fusion protein as a strategy to improve the immunogenicity of GRA1 and to construct the cocktail vaccine.Methods PCR technique was used to amplify HBsAg and GRA1 respectively. The PCR products were digested by restriction enzymes BclⅠ/EcoRⅠfor HBsAg and EcoRⅠ/XhoⅠ for GRA1 and subcloned into pcDNA3 BamHⅠand XhoⅠsites.The recombinant(pcDNA3-HBsAg- GRA1) was identified with appropriate restriction enzymes and DNA sequencing analysis.Results the chimeric pcDNA3-HBsAg-GRA1plasmid have been constructed successfully.Conclusion The result will lay the foundation for further study on the protective immunity of pcDNA3-HBsAg-GRA1against T. gondii or hepatitis B virus infection.
出处
《中国人兽共患病杂志》
CSCD
北大核心
2004年第2期122-125,共4页
Chinese Journal of Zoonoses
基金
湖南省"十五"重点学科建设项目资助
