Tissue specificity of the initiation of immunoglobulin kappa gene transcription.

F G Falkner, E Neumann, H G Zachau
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引用次数: 34

Abstract

The transient transcription of a rearranged mouse immunoglobulin kappa gene was studied in a monkey fibroblast cell line. The gene was inserted into an SV40 expression vector and the calcium phosphate coprecipitation method was used for transfection. The transcripts were correctly spliced; transcription, however, was initiated within the vector and not at the correct site 23-26 bp upstream of the gene, irrespective of the length of the upstream sequences (90, 160, 370, and 870 bp) in the plasmid constructs. In contrast, accurately initiated transcripts were observed when a plasmid containing the kappa gene with 870 bp of its upstream sequence was introduced into a lymphoid cell line; the plasmid was constructed from the pSV2-gpt vector and the electric impulse method was used for gene transfer in most experiments. Tissue-specific expression of kappa light chain genes in lymphoid cells is known to depend on the presence of an enhancer element in the J-C intron. The results reported in this paper suggest that the sequence elements pd and dc which are located upstream of the leader gene segment also act in a tissue-specific manner and that it is the initiation of transcription which is a tissue-specific event.

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免疫球蛋白kappa基因转录起始的组织特异性。
在猴成纤维细胞系中研究了重排小鼠免疫球蛋白kappa基因的瞬时转录。将该基因插入SV40表达载体,采用磷酸钙共沉淀法转染。转录本正确拼接;然而,无论质粒结构中上游序列(90、160、370和870 bp)的长度如何,转录都是在载体内开始的,而不是在基因上游23-26 bp的正确位点。相比之下,将含有kappa基因上游序列870 bp的质粒导入淋巴样细胞系,可以准确地启动转录物;质粒以pSV2-gpt为载体构建,多数实验采用电脉冲法进行基因转移。已知kappa轻链基因在淋巴样细胞中的组织特异性表达依赖于J-C内含子中增强子的存在。本文的研究结果表明,位于先导基因片段上游的序列元件pd和dc也具有组织特异性,是组织特异性的转录起始事件。
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