Transfection and recombination with molecularly cloned derivatives of avian sarcoma virus UR2.
Transfection and recombination with molecularly cloned derivatives of avian sarcoma virus UR2.
复制标题
禽肉瘤病毒 UR2 分子克隆衍生物的转染和重组。
DOI：
10.1016/0042-6822(86)90469-1
复制
发表时间：
1986
期刊：
影响因子：
3.7
通讯作者：
Balduzzi,PC
中科院分区：
文献类型：
--
作者：
Das,KS;Christensen,JR;Balduzzi,PC
已经制备了禽肉瘤病毒UR 2的克隆版本，质粒pKD 6，其包括两个LTR区域之间的完整的非置换前病毒序列。质粒在转染实验中具有生物活性，即使是完整的。从pKD 6中构建了两个转化缺陷型突变体，它们在转化基因中具有非重叠缺失。当两者的混合DNA与辅助病毒DNA一起用于转染鸡胚成纤维细胞沿着时，这些突变体重组产生转化病毒。然而，重组不容易检测到细胞时，共感染的液体收获的文化分别转染DNA从每个突变体。这一点，和标志物救援实验与温度敏感突变体的UR2的转化缺陷，但能够复制，表明删除突变体的UR2不繁殖有效。
A cloned version of avian sarcoma virus UR2, plasmid pKD6, which includes the full, nonpermuted proviral sequence between two LTR regions, has been prepared. The plasmid is biologically active in transfection experiments, even when intact. Two transformation-defective mutants with nonoverlapping deletions within the transforming generoswere constructed from pKD6. These mutants recombine to produce transforming virus when mixed DNA from both is used to transfect chick embryo fibroblasts along with helper virus DNA. However, recombination was not readily detected when cells were coinfected with fluids harvested from cultures separately transfected with DNA from each mutant. This, and marker rescue experiments with a temperature-sensitive mutant of UR2 defective in transformation but able to replicate, suggest that deletion mutants of UR2 do not propagate efficiently.