摘要
采用2种不同的逆转录病毒载体系统(pMX和pMSCV),构建携带绿色荧光蛋白(EGFP)基因的重组载体,探索能高效感染水牛胎儿成纤维细胞(BFFs)的病毒系统和感染方法.结果发现,2种逆转录病毒系统包装出的重组病毒滴度都能达到106,将pMX病毒系统生产的重组病毒经超速离心浓缩后,滴度可达到107;2种重组病毒都能感染BFFs,但pMX系统比pMSCV系统更能有效地感染BFFs;通过对病毒的感染方式进行比较,发现使用新鲜病毒连续感染BFFs 2次(每次间隔12 h),或者经过超速离心浓缩后的病毒感染BFFs 1次,可以显著提高感染效率.
After two retroviral systems with enhanced green fluorescent protein （ EGFP） report gene were constructed in this study , the retroviral systems and the infection method suitable for buffalo fetal fibro-blasts （BFFs） were investigated.The results revealed that the retrovirus titer of the two systems was 106, and the titer of the pMX system could be up to 10 7 after ultracentrifugation concentration .The infection efficiency of pMX retroviral system was higher than that of the pMSCV retroviral system .Using the fresh retrovirus infected BFFs twice （12 h interval） or ultracentrifugal retrovirus infected BFFs once could im-prove the infection efficiency .
出处
《华南农业大学学报》
CAS
CSCD
北大核心
2013年第4期558-563,共6页
Journal of South China Agricultural University
基金
863计划项目(2011AA100607)
国家自然科学基金(30960251)
广西教育厅资助项目(200911MS12)
广西大学科研基金项目(XBZ120572)
