RETROTRANSPOSITION OF A MOUSE IAP SEQUENCE TAGGED WITH AN INDICATOR GENE
RETROTRANSPOSITION OF A MOUSE IAP SEQUENCE TAGGED WITH AN INDICATOR GENE
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DOI：
10.1016/0092-8674(91)90217-m
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发表时间：
1991-01-11
期刊：
影响因子：
64.5
通讯作者：
HEIDMANN, T
中科院分区：
文献类型：
--
作者：
HEIDMANN, O;HEIDMANN, T
我们用含有新霉素的指示基因标记了克隆的小鼠 IAP 序列，该指示基因的表达受转座子通过 RNA 中间体的影响。引入肿瘤细胞的标记IAP的转座可以通过简单选择G418中的细胞来检测，频率为每代每个细胞10(-6)。 PCR 扩增后的 Southern 印迹分析和核苷酸测序证明了标记元件的“逆转录转座”，其中包含指示基因中包含的内含子的剪接，以及转座 IAP 的逆转录病毒样逆转录和整合，以及已识别靶位点的 6 bp 重复。发现转座对于该元件具有诱变性，如果所识别的标记和内源 IAP 转录物以二聚体的形式共壳到 IAP 颗粒中，则可能会出现这种情况。
We have marked a cloned mouse IAP sequence with a neomycin-containing indicator gene whose expression is conditioned by passage of the transposon through an RNA intermediate. Transposition of the marked IAP introduced into tumor cells could be detected by simple selection of the cell in G418, at a frequency of 10(-6) per cell per generation. Southern blot analysis and nucleotide sequencing after PCR amplification demonstrated "retrotransposition" of the marked element, with splicing out of an intron contained in the indicator gene, and retroviral-like reverse transcription and integration of the transposed IAPs, with 6 bp duplications of the identified target sites. Transposition was found to be mutagenic for the element, as might be expected if the identified marked and endogenous IAP transcripts were coencapsidated into IAP particles as dimers.