Characterization of a stable eukaryotic cell line expressing the Rous sarcoma virus integrase.
Characterization of a stable eukaryotic cell line expressing the Rous sarcoma virus integrase.
复制标题
表达劳斯肉瘤病毒整合酶的稳定真核细胞系的表征。
DOI：
10.1016/0042-6822(92)90574-9
复制
发表时间：
1992
期刊：
影响因子：
3.7
通讯作者：
Grandgenett,DP
中科院分区：
文献类型：
--
作者：
Mumm,SR;Hippenmeyer,PJ;Grandgenett,DP
劳斯肉瘤病毒整合蛋白（IN）是病毒DNA有效整合到宿主基因组中所必需的。使用牛乳头瘤病毒载体在小鼠C127细胞中表达IN。该系统利用小鼠金属硫蛋白启动子和SV 40晚期多聚腺苷酸化信号有效表达IN。一个稳定的细胞系来源于一个单一的潮霉素耐药菌落进行了表征。IN的表达增加显着锌+诱导的金属硫蛋白启动子，但没有响应“超诱导”协议。全长非磷酸化IN是表达的主要产物。还产生了由内部Met密码子处的翻译起始产生的次要产物。所表达的IN在其COOH-末端没有表现出多肽异质性，也没有如IN从病毒体免疫沉淀时所见的磷酸化。使用亚细胞分级分离和间接免疫荧光，IN主要定位于细胞核，在一些细胞中似乎集中在细胞核内的离散位点。
The Rous sarcoma virus integration protein (IN) is required for efficient integration of viral DNA into the host genome. IN was expressed in mouse C127 cells using a bovine papillomavinus vector. This system utilizes the mouse metallothionein promoter and the SV40 late polyadenylation signal for efficient expression of IN. A stable cell line derived from a single hygromycin-resistant colony was characterized. The expression of IN increased significantly upon Zn2+induction of the metallothionein promoter, but did not respond to “superinduction” protocols. Full-length nonphosphorylated IN was the major product of expression. A minor product resulting from initiation of translation at an internal Met codon was also produced. The expressed IN did not exhibit the polypeptide heterogeneity at its COOH-terminus nor phosphorylation as is seen when IN is immunoprecipitated from virions. Using subcellular fractionation and indirect immunofluorescence, IN was primarily localized to nuclei and in some cells appeared to concentrate at discrete loci within the nuclei.