Induction of human beta-interferon synthesis with poly(rI . rC) in mouse cells transfected with cloned cDNA plasmids.
Induction of human beta-interferon synthesis with poly(rI . rC) in mouse cells transfected with cloned cDNA plasmids.
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在用克隆的 cDNA 质粒转染的小鼠细胞中用聚 (rI.rC) 诱导人 β-干扰素合成。
DOI：
10.1073/pnas.79.14.4337
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发表时间：
1982
影响因子：
11.1
通讯作者：
Hayward,GS
中科院分区：
文献类型：
--
作者：
Pitha,PM;Ciufo,DM;Kellum,M;Raj,NB;Reyes,GR;Hayward,GS
通过与单纯疱疹病毒胸苷激酶（TK）基因共转染，将人基因组DNA和携带人β-干扰素cDNA基因部分的质粒导入小鼠Ltk-细胞。一个质粒含有与插入到pBR 322中的前-β-干扰素mRNA互补的840个碱基对的人DNA，而其它质粒具有仅含有在TK启动子的转录控制下插入的560个碱基对的编码区的杂合基因。在任何检测的小鼠TK+细胞系中均未检测到组成型干扰素产生。然而，干扰素的合成可以被poly（rI）诱导。rC）在至少16个这些细胞系中处理，包括用基因组DNA、β-干扰素cDNA和TK-β-干扰素cDNA杂合基因转染的克隆。所产生的干扰素对人细胞具有特异性，并且可以被抗人β-干扰素的抗血清中和。与其中诱导的β-干扰素的合成是瞬时的人成纤维细胞相反，聚（rI . rC）诱导的TK+细胞系持续产生β-干扰素，持续时间延长，并且对超诱导条件没有反应。因此，在转染的小鼠细胞中，来自人β-干扰素基因的编码DNA序列，没有任何相邻的3'或5'侧翼的人DNA序列，足以指导生物活性产物的合成和响应在人细胞中操作的特异性诱导系统。然而，在人细胞中关闭诱导干扰素合成的机制似乎在用β-干扰素cDNA转染的小鼠细胞中不起作用。
Human genomic DNA and plasmids carrying portions of the cDNA gene for human beta-interferon have been introduced into mouse Ltk- cells by cotransfection with a herpes simplex virus thymidine kinase (TK) gene. One plasmid contains 840 base pairs of human DNA complementary to pre-beta-interferon mRNA inserted into pBR322, whereas the other plasmids have hybrid genes containing only the 560-base pair coding region inserted under the transcriptional control of the TK promoter. Constitutive interferon production could not be detected in any of the mouse TK+ cell lines tested. Nevertheless, synthesis of interferon could be induced by poly(rI . rC) treatment in at least 16 of these cell lines, including clones transfected with genomic DNA, the beta-interferon cDNA, and the TK-beta-interferon cDNA hybrid gene. The interferon produced was specific for human cells and could be neutralized by antiserum against human beta-interferon. In contrast to human fibroblast cells, in which the synthesis of induced beta-interferon is transient, the poly(rI . rC)-induced TK+ lines continued to produce beta-interferon for prolonged periods of time and did not respond to superinduction conditions. Therefore, in transfected mouse cells, the coding DNA sequence from the human beta-interferon gene, without any of the adjacent 3' or 5' flanking human DNA sequences, was sufficient both to direct synthesis of biologically active product and to respond to the specific induction system that operates in human cells. However, the mechanism that switches off the synthesis of induced interferon in human cells appears not to operate in mouse cells transfected with beta-interferon cDNA.