Falkner F G, Neumann E, Zachau H G
Hoppe Seylers Z Physiol Chem. 1984 Nov;365(11):1331-43. doi: 10.1515/bchm2.1984.365.2.1331.
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.
在猴成纤维细胞系中研究了重排的小鼠免疫球蛋白κ基因的瞬时转录。将该基因插入SV40表达载体,并采用磷酸钙共沉淀法进行转染。转录本被正确剪接;然而,转录起始于载体内部,而非基因上游23 - 26 bp的正确位点,无论质粒构建体中上游序列(90、160、370和870 bp)的长度如何。相反,当将含有κ基因及其870 bp上游序列的质粒导入淋巴样细胞系时,观察到了准确起始的转录本;该质粒由pSV2 - gpt载体构建而成,在大多数实验中采用电脉冲法进行基因转移。已知淋巴样细胞中κ轻链基因的组织特异性表达依赖于J - C内含子中增强子元件的存在。本文报道的结果表明,位于前导基因片段上游的序列元件pd和dc也以组织特异性方式发挥作用,并且转录起始是一个组织特异性事件。