重组逆转录病毒的制备、浓缩及保存研究

Studies on vitro preparation of recombinant retrovirus, concentrate and storage

选择含有新霉素磷酸转移酶Ⅱ（ＮｅｏＲ）的重组逆转录病毒载体，用包装细胞进行包装，得到了含有重组逆转录病毒粒子的病毒上清，该病毒上清经差速离心法浓缩后，－８０℃无任何添加物的情况下保存３个月，体外感染靶细胞ＮＩＨ３Ｔ３以测定其滴度，冻存前后的浓缩病毒上清的感染滴度无明显降低，经过冻存的浓缩病毒上清的感染滴度仍可达到１０６ｃｆｕ／ｍｌ，具有理想的感染靶细胞的能力。

Recombinant retroviral vector(pLXSN) carrying NeoR were transfected into PA3 17 by lipofectin and calcium phosphate precipitation. The cell supernatants from G418- resistant PA317 in which there are recombinant retroviral were concentrated through centrifuge at different speeds at 4℃. The concen-trated viral supernatants were stored at- 80℃ for three months. NIH3T3 were infected to measure the titer of viral supernatants. The results showed there are no marked differences between the titer of concentrated viral supernatants and unconcentrated viral supernatants, the titer of concentrated viral supernatants stored at- 80℃ for three months can still attain 106 cfu/ml, and it has the ideal capacity of infecting target cell.