Kuff E L, Feenstra A, Lueders K, Smith L, Hawley R, Hozumi N, Shulman M
Proc Natl Acad Sci U S A. 1983 Apr;80(7):1992-6. doi: 10.1073/pnas.80.7.1992.
We analyzed two functionally defective mouse kappa light chain gene variants previously shown to contain novel insertions of repetitive DNA in their intervening sequences [Hawley, R. G., Shulman, M. J., Murialdo, H., Gibson, D. M. & Hozumi, N. (1982) Proc. Natl. Acad. Sci. USA 79, 7425-7429]. Heteroduplex analysis of the cloned genes shows that the insertions consist of intracisternal A-particle (IAP) genetic elements. Each insertion includes an IAP 5' long terminal repeat (LTR) sequence and extends to a characteristic IAP internal BamHI site where the IAP sequence is interrupted because the mutant genes were cloned from complete BamHI digests of the cellular DNAs. Restriction enzyme mapping indicates that the 5' LTR boundaries of the inserted IAP elements correspond closely to the previously determined rearrangement sites in the mutant genes. The IAP insertions in the two mutants can be distinguished by restriction-site differences and by the fact that one of them contains a deletion that is absent in the other. Nucleotide sequence data are presented for the LTRs of one full-length IAP gene copy randomly selected from a mouse genomic DNA library. These LTRs show many features typical of known integrated retroviral terminal repeat units, and the entire gene is bracketed by short direct repeats within the adjacent cellular DNA. Thus, the findings show that IAP genetic elements can appear in new locations in mouse cellular DNA and suggest that this may occur through a process of proviral insertion.
我们分析了两个功能缺陷型小鼠κ轻链基因变体，之前已表明它们在间隔序列中含有重复DNA的新插入片段[霍利，R.G.，舒尔曼，M.J.，穆里亚尔多，H.，吉布森，D.M.和小泉，N.（1982年）《美国国家科学院院刊》79，7425 - 7429]。对克隆基因的异源双链分析表明，插入片段由核内A颗粒（IAP）遗传元件组成。每个插入片段都包含一个IAP 5'长末端重复（LTR）序列，并延伸至一个特征性的IAP内部BamHI位点，在该位点IAP序列被打断，因为突变基因是从细胞DNA的完整BamHI酶切片段中克隆得到的。限制性酶切图谱分析表明，插入的IAP元件的5' LTR边界与突变基因中先前确定的重排位点紧密对应。两个突变体中的IAP插入片段可通过限制性位点差异以及其中一个含有另一个所没有的缺失这一事实来区分。本文给出了从一个小鼠基因组DNA文库中随机选取的一个全长IAP基因拷贝的LTR的核苷酸序列数据。这些LTR显示出许多已知整合逆转录病毒末端重复单元的典型特征，并且整个基因被相邻细胞DNA中的短直接重复序列所包围。因此，这些发现表明IAP遗传元件可以出现在小鼠细胞DNA的新位置，并提示这可能是通过前病毒插入过程发生的。
