摘要
目的构建以结核分枝杆菌热休克蛋白65和汉坦病毒G2重组融合基因为基础的核酸疫苗。方法采用聚合酶链反应从结核杆菌H37Rv株基因中,扩增出HSP65的编码基因,从T-H8205G2质粒扩增出G2蛋白的编码基因,经相应限制性核酸内切酶消化后,插入真核表达载体pcDNA3·1/His,并将此重组质粒通过酶切、SDS-PAGE分别测定表达产物的相对分子量。结果获得正向插入的pcDNA3·1/His-HSP65-G2重组表达质粒,与理论预期大小一致。结论以编码HSP65和G2抗原基因的重组基因的真核表达载体构建成功,为进一步研究其在结核病和出血热防治中的作用奠定了基础。
Objective To construct nucleotide vaccine based on the recombinant plasmids of hsp65 gene of mycobacterium tuberculosis H37Rv strain and G2 gene fragments of Hantavirus H8205 slxain. Methods The gene encoding heat shock protein 65 kd and G2 glycoprotein were amplified from genome of Mycobacterium tuberculosis and T - H8205G2 plasmid by routine polymerase chain reaction （PCR）. After corresponding to restriction endonuclease digestion, the fragment were inserted into downstream of the vector pCDNA3.1/His to obtain the recombinant eukaryotic expression plasmid poDNA3.1/His - hsp65 - G2. The recombinant plasmid pcDNA3.1/His - hsp65 - G2 were used to analyze the molecular weight respectively. Results The positive recombinant plasmid harbouring the right inserted G2 glyeoprotein gene and hsp65 gene fragment was obtained. Conclusion The successful construction of recombinant eukaryotic expression vector based on hsp65 gene of mycobacterium tuberculosis and G2 gene fragments of Hantavirus H8205 staain ,lays the foundation for further researching in prevention and treatment of tuberculosis and HFRS.
出处
《中国热带医学》
CAS
2006年第9期1541-1542,1560,共3页
China Tropical Medicine
基金
武汉市科技攻关计划项目(NO2002600513415)
