Gene-transfer, stability, and biochemical properties of animal cells transformed with vaccinia DNA.
Gene-transfer, stability, and biochemical properties of animal cells transformed with vaccinia DNA.
复制标题
用痘苗 DNA 转化的动物细胞的基因转移、稳定性和生化特性。
DOI：
10.1016/0042-6822(82)90236-7
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发表时间：
1982
期刊：
影响因子：
3.7
通讯作者：
Esteban,M
中科院分区：
文献类型：
--
作者：
Pellicer,A;Esteban,M
通过基因转移技术将牛痘病毒的大DNA（180 kb）导入动物细胞并维持。虽然在一些克隆中可以发现所有或几乎所有的牛痘基因组，但没有观察到细胞病变效应，表明牛痘DNA是非感染性分子。此外，牛痘病毒胸苷激酶（tk）基因不能补充小鼠LtK（−）细胞的突变表型。通过Southern印迹分析转化细胞中病毒序列的排列，其中克隆保持在培养物中50至200代细胞。一些病毒序列似乎被整合到高分子量细胞DNA中。用克隆的牛痘病毒DNA末端片段作探针，我们获得了许多转化子中存在游离病毒DNA分子的证据。分析了这些转化体的许多生物学特性。最显著的观察结果是这些细胞不能支持外源添加的牛痘病毒的复制，而VSV和Mengo病毒的复制不受影响。这种干扰样现象与转化细胞中病毒序列的存在相关。
The introduction and maintenance of the large DNA (180 kb) from vaccinia virus into animal cells has been accomplished by gene transfer techniques. While all or nearly all vaccinia genome could be found in some clones, no cytopathic effects were observed, indicating that vaccinia DNA was a noninfectious molecule. In addition, the vaccinia virus thymidine kinase (tk) gene was unable to complement the mutant phenotype of mouse LtK(−) cells. The arrangement of viral sequences in transformed cells was analyzed by Southern blots with clones maintained in culture from 50 to 200 cell generations. Some of the viral sequences appear to be integrated into high-molecular-weight cellular DNA. Using the cloned terminal fragment of vaccinia DNA as probe, we have obtained evidence which indicates that free viral DNA molecules are present in a number of transformants. A number of biological properties of these transformants was analyzed. The most significant observation was the failure of these cells to support the replication of exogenously added vaccinia virus while the replication of VSV and Mengo viruses was unaffected. This interference-like phenomenon is correlated with the presence of viral sequences in the transformed cells.