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首页> 外文期刊>Molecular and Cellular Biology >Inducible expression of the human interferon beta 1 gene linked to a bovine papilloma virus DNA vector and maintained extrachromosomally in mouse cells.
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Inducible expression of the human interferon beta 1 gene linked to a bovine papilloma virus DNA vector and maintained extrachromosomally in mouse cells.

机译：与牛乳头瘤病毒DNA载体连接的人干扰素β1基因的可诱导表达，并在小鼠细胞内保持染色体外。

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摘要

A 1.6-kilobase DNA segment of the genomic human interferon beta 1 (IF-beta 1) gene was inserted into each of two possible orientations at the single HindIII site of a recombinant plasmid pBPV69T, consisting of the 69% transforming region of the bovine papilloma virus type 1 (BPV-1) and a modified SalI-SalI fragment of plasmid pBR322. After cleavage of the pBR322 sequences from this recombinant, BPV69T-IF-beta 1 hybrid DNAs were transfected onto C127 mouse cells by the standard calcium precipitation technique. Mouse cells transformed by this hybrid DNA produced low levels of human IF-beta 1 constitutively and responded to induction with either inactivated Newcastle disease virus or polyriboinosinic acid-polyribocytidylic acid. The BPV69T-IF-beta 1 hybrid DNA was nonintegrated in the transformed mouse cells but had acquired DNA sequences as a result of the transfection. Accurate transcripts of the IF-beta 1 mRNA were detected in cells only after induction. When the IF-beta 1 gene was oriented in the plasmid in the same direction of transcription as the BPV-1 genome, transcription was promoted from within the BPV-1 sequences. These results indicate that the regulatory sequences responsible for the inducible expression of the human IF-beta 1 gene are present in the 1.6-kilobase genomic segment and that these sequences can function in a free extrachromosomal state linked to BPV-1 sequences.
机译：将基因组人干扰素beta 1（IF-beta 1）基因的1.6碱基碱基DNA片段插入重组质粒pBPV69T的单个HindIII位点的两个可能方向的每个方向，该位置由牛乳头状瘤的69％转化区组成病毒1型（BPV-1）和修饰的质粒pBR322的SalI-SalI片段。从该重组体切割pBR322序列后，通过标准钙沉淀技术将BPV69T-IF-β1杂合DNA转染到C127小鼠细胞上。由这种杂合DNA转化的小鼠细胞组成型地产生了低水平的人IF-β1，并响应了灭活的新城疫病毒或多核糖肌酸-多核糖酸的诱导。 BPV69T-IF-β1杂合DNA未整合到转化的小鼠细胞中，但由于转染而获得了DNA序列。仅在诱导后才在细胞中检测到IF-β1mRNA的准确转录本。当IF-beta 1基因在质粒中的转录方向与BPV-1基因组相同时，则从BPV-1序列内部促进转录。这些结果表明负责人IF-β1基因的诱导表达的调控序列存在于1.6碱基对的基因组片段中，并且这些序列可以在与BPV-1序列连接的游离染色体外状态下发挥作用。

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