Gilmore-Herbert M, Hercules K, Komaromy M, Wall R
Proc Natl Acad Sci U S A. 1978 Dec;75(12):6044-8. doi: 10.1073/pnas.75.12.6044.
UV transcription mapping with recombinant DNA probes containing immunoglobulin kappa light chain mRNA sequences has been used to determine the size of the transcription unit coding for kappa light chain m RNA and to establish the arrangement of variable and constant regions in this transcription unit. In relation to ribosomal RNA standards, the transcription of kappa light chain constant region sequences into nuclear RNA exhibits a UV target size of 9.6 kbases (kb). The kappa light chain variable region exhibits a UV target size of 7.6 kb indicating that it is separated by approximately 2.0 kb from the constant region in the kappa light chain transcription unit. The size of the primary transcript (i.e., the direct, unprocessed RNA product of transcription) predicted from the constant region target size concurs with our previous pulse-labeling results which showed that the largest presumptive nuclear RNA precursor to kappa light chain mRNA is approximately 10 kb. In addition, the UV target size of cytoplasmic kappa mRNA is indistinguishable from the target size of constant region sequences in nuclear RNA. These results suggest that the kappa light chain transcription unit is copied directly into a 10-kb nuclear RNA precursor in which the kappa variable and constant regions are separated by approximately 2 kb. Accordingly, it is proposed that the joining of immunoglobulin kappa light chain variable and constant regions occurs in the post-transcriptional processing of this large nuclear RNA precursor into kappa light chain mRNA.
利用含有免疫球蛋白κ轻链mRNA序列的重组DNA探针进行紫外线转录图谱分析,已用于确定编码κ轻链mRNA的转录单位的大小,并确定该转录单位中可变区和恒定区的排列方式。相对于核糖体RNA标准,κ轻链恒定区序列转录成核RNA时,紫外线靶标大小为9.6千碱基(kb)。κ轻链可变区的紫外线靶标大小为7.6 kb,表明在κ轻链转录单位中,它与恒定区相隔约2.0 kb。根据恒定区靶标大小预测的初级转录本(即转录的直接、未加工的RNA产物)的大小与我们之前的脉冲标记结果一致,该结果表明,κ轻链mRNA最大的假定核RNA前体约为10 kb。此外,细胞质κ mRNA的紫外线靶标大小与核RNA中恒定区序列的靶标大小无法区分。这些结果表明,κ轻链转录单位直接转录成一个10 kb的核RNA前体,其中κ可变区和恒定区相隔约2 kb。因此,有人提出免疫球蛋白κ轻链可变区和恒定区的连接发生在这个大的核RNA前体转录后加工成κ轻链mRNA的过程中。