Differential transient and long-term expression of DNA sequences introduced into T-lymphocyte lines.
Differential transient and long-term expression of DNA sequences introduced into T-lymphocyte lines.
复制标题
引入 T 淋巴细胞系的 DNA 序列的差异瞬时和长期表达。
DOI：
10.1089/dna.1.1986.5.439
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Rothenberg,EV
中科院分区：
文献类型：
--
作者：
Novak,TJ;Rothenberg,EV
我们利用原生质体融合技术将编码新霉素磷酸转移酶（neo）和氯霉素乙酰转移酶（CAT）的基因导入小鼠和人t淋巴细胞系。在Rous肉瘤病毒长末端重复序列（RSV LTR）、SV40早期区或单纯疱疹病毒胸苷激酶基因（HSV TK）启动子的控制下，含有该基因的质粒构建可以稳定地将三种t细胞系转化为G-418抗性。不同细胞系的特征频率可以相差至少两个数量级，尽管初始DNA摄取和瞬时表达是相似的。在这两种小鼠细胞系中，长期转化体中保留了少量的基因拷贝。在整合之前，对癌新基因产物的瞬时表达分析表明，不同结构的内在启动子强度的差异进一步受到被转录的编码序列的影响。因此，虽然neo蛋白在Rous LTR和SV40早期启动子中的瞬时表达是相似的，但Rous LTR对CAT蛋白合成的指导水平比SV40早期启动子高两个数量级。
We have used a protoplast fusion protocol to introduce the genes encoding neomycin phosphotransferase (neo) and chloramphenicol acetyltransferase (CAT) into murine and human T-lymphocyte lines. Plasmid constructs containing theneogene under the control of the promoters from the Rous sarcoma virus long terminal repeat (RSV LTR), the SV40 early region, or the herpes simplex virus thymidine kinase gene (HSV TK) can stably transform each of three T-cell lines to G-418 resistance. The characteristic frequencies for different cell lines can differ by at least two orders of magnitude, although initial DNA uptake and transient expression are similar. In the two murine cell lines, low numbers of gene copies are retained in long-term transformants. Prior to integration, transient expression assays forcatorneogene products reveal that the differences in intrinsic promoter strength of different constructs are further influenced by the coding sequences being transcribed. Thus, while transient expression of the neo protein is similar from both the Rous LTR and the SV40 early promoter, the Rous LTR directs synthesis of CAT protein at levels two orders of magnitude higher than those from the SV40 early promoter.