Gruss P, Khoury G
Proc Natl Acad Sci U S A. 1981 Jan;78(1):133-7. doi: 10.1073/pnas.78.1.133.
The complete rat preproinsulin gene I was cloned into a simian virus 30 (SV 40) vector. Most of the late region of the viral vector, including the SV40 intervening sequences (introns) and all of the major splice junctions, was deleted and replaced by the entire rat insulin gene. The recombinant molecules and a temperature-sensitive helper virus (tsA28) were inoculated into monkey kidney cultures. The formation of stable transcripts of the insulin insert was as efficient as the production of late SV40 mRNA. Analysis of these transcripts indicated that the rat preproinsulin gene nucleotide signals involved in RNA splicing and poly(A) addition were used. Examination of the 5' ends of the mRNAs showed several classes, one of which was the same size as the authentic rat insulinoma mRNA. This suggests that a portion of the transcripts may be initiated or processed faithfully, or both, at their 5' ends within rat insulin sequences. Significant quantities of a protein identified as rat proinsulin were synthesized. Detection of most of the proinsulin in the tissue culture medium suggests that this protein was secreted.
完整的大鼠胰岛素原基因I被克隆到猿猴病毒30（SV40）载体中。病毒载体的大部分晚期区域，包括SV40间隔序列（内含子）和所有主要剪接位点，均被删除，取而代之的是完整的大鼠胰岛素基因。将重组分子和一种温度敏感型辅助病毒（tsA28）接种到猴肾培养物中。胰岛素插入片段稳定转录本的形成与晚期SV40 mRNA的产生效率相同。对这些转录本的分析表明，大鼠胰岛素原基因中参与RNA剪接和聚腺苷酸化的核苷酸信号被利用。对mRNA 5'末端的检查显示出几类，其中一类与真正的大鼠胰岛素瘤mRNA大小相同。这表明一部分转录本可能在其5'末端的大鼠胰岛素序列内被正确起始或加工，或两者兼而有之。合成了大量被鉴定为大鼠胰岛素原的蛋白质。在组织培养基中检测到大部分胰岛素原，表明该蛋白质被分泌了。
