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.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2008年第1期16-19,共4页
Chinese Journal of Microbiology and Immunology
关键词
实时定量PCR
重组痘苗病毒
传代
稳定性
Real-time quantitative PCR
Recombinant vaccinia
Passage
Stability
