摘要
对结核病核酸疫苗的质粒DNA的制备性纯化过程进行了初步研究。此工艺主要包括以简化的碱裂解法处理大肠杆菌、超滤与离心法除大部分杂质并浓缩样品、Q-Sepharose阴离子交换层析、DNA B ioSepharTMFF亲和层析等步骤,以去除细胞RNA和蛋白等杂质,并对不同分子结构的质粒DNA作分离。结果表明:以碱裂液中质粒DNA为计算基准的得率为56.61%、超螺旋质粒DNA的电泳纯度100%。
The purification process for the production of tuberculosis DNA vaccine was studied. The plasmids of DNA vaccine（pVAX-44） in Escherichia coli（DHSa） were separated and purified by the simplified method of alkaline lysis, ultra-filtration, centrifuge, and Q-Sepharose anion exchange chromatography. The plasmid DNA molecules in the supercoiled form were separated from other DNA molecules through DNA BioSepharTM fast flow affinity chromatography. The final yield of plasmid DNA from the broth after alkaline lysis is 56.61%, and its purity determined by agarose electrophoresis is almost 100%.
出处
《华东理工大学学报（自然科学版）》
EI
CAS
CSCD
北大核心
2006年第11期1285-1288,共4页
Journal of East China University of Science and Technology
关键词
结核病
核酸疫苗
质粒
纯化
层析
tuberculosis
DNA vaccine
plasmid
purification
chromatography
