不同逆转录载体系统应用于水牛胎儿成纤维细胞转基因的探索

Infection of Buffalo Fetal Fibroblasts by Two Retrovirus Systems

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.

Abstract: 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 fibroblasts (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 10⁷ 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 improve the infection efficiency.