实时定量PCR方法监测重组痘苗外源基因传代稳定性  

Real-time PCR assay to monitor the stability of exterior gene in different recombinant vaccinia passages

作　　者：王宁[1] 李媛媛[1] 徐静[1] 王芙[1] 戴凯凡[2] 刘颖[2] 罗兰[1] 邵一鸣 刘建源[1] 

机构地区：[1]北京生物制品研究所免疫研究室,100024 [2]中国疾病预防控制中心性病艾滋病预防控制中心

出　　处：《中华微生物学和免疫学杂志》2008年第1期16-19,共4页Chinese Journal of Microbiology and Immunology

摘　　要：目的建立实时定量PCR方法以监测外源基因在不同重组痘苗代次中的稳定性。方法裂解法提取8个代次插入有HIV基因的重组痘苗DNA，用实时定量PCR的绝对和相对定量方法，研究不同代次基因组中插入的外源基因与痘苗基因组的数量关系。结果绝对和相对定量方法均证明，在传代过程中，重组痘苗中的外源基因有缺失现象。结论建立的实时定量PCR方法可以特异、定量地分析外源基因在重组痘苗中的稳定性。Objective To establish a real-time quantitative PCR method to analyze the stability of exterior gene in different recombinant virus passages. Methods The DNA of eight passages of recombinant vaccinia viruses containing HIV gene were extracted, and analyzed by absolute or relative real-time quantitative PCR. Then the ratio of the HIV gene copies to vaccinia genome copies was determined. Results Abso- lute and relative real-time quantitative PCR assay result both suggest that the HIV gene was lost during recombinant vaccinia viruses passages. Conclusion The novel specific real-time quantitative PCR assay to analyze the stability of exterior gene in vaccinia virus genome has been developed.

关 键 词：实时定量PCR 重组痘苗病毒 传代 稳定性 

分 类 号：R686[医药卫生—骨科学]