摘要
选择含有新霉素磷酸转移酶Ⅱ (Neor)的重组逆转录病毒载体 ,通过包装细胞进行包装 ,得到了含有重组逆转录病毒粒子的病毒溶液 ,该病毒溶液分别采用差速离心和滤膜截留两种方法进行浓缩 ,浓缩前、后的病毒溶液体外感染靶细胞NIH3T3以测定其滴度。结果显示 :差速离心法的浓缩效率要优于滤膜截留法的浓缩效率 ,其浓缩效率能达到 34 5%。
Recombinant retroviral vectors(pLXSN) carrying Neo r were transfected into PA317 by lipofectin and calcium phosphate coprecipitation.The cell supernatants from G418-resistant PA317 in which there are recombinant retrovirus were concentrated through speed differential centrifugation and membrane filteration,respectively,NIH3T3 cells were infected by the concentrated virus to measure its titer.The results showed that the concentration efficiency of speed differential centrifugation is superior to that of membrane filteration and the concentration efficiency of speed differential centrifugation speed can attain 34.5%.
出处
《生物学杂志》
CAS
CSCD
2001年第2期11-13,共3页
Journal of Biology
基金
国家"8 63"计划资助项目!(Z2 1- 0 4- 0 3)
关键词
重组逆转录病毒
差速离心
滤膜截留
浓缩方法
载体
recombinant retrovirus
concentration
speed differential centrifugation
membrane filteration.
