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通过P因子诱导的缺口修复在果蝇中进行靶向基因替换。

Targeted gene replacement in Drosophila via P element-induced gap repair.

作者信息

Gloor G B, Nassif N A, Johnson-Schlitz D M, Preston C R, Engels W R

机构信息

Faculty of Medicine, Memorial University of Newfoundland, St. John's, Canada.

出版信息

Science. 1991 Sep 6;253(5024):1110-7. doi: 10.1126/science.1653452.

Abstract

Transposable elements of the P family in Drosophila are thought to transpose by a cut-and-paste process that leaves a double-strand gap. The repair of such gaps resulted in the transfer of up to several kilobase pairs of information from a homologous template sequence to the site of P element excision by a process similar to gene conversion. The template was an in vitro-modified sequence that was tested at various genomic positions. Characterization of 123 conversion tracts provided a detailed description of their length and distribution. Most events were continuous conversion tracts that overlapped the P insertion site without concomitant conversion of the template. The average conversion tract was 1379 base pairs, and the distribution of tract lengths fit a simple model of gap enlargement. The conversion events occurred at sufficiently high frequencies to form the basis of an efficient means of directed gene replacement.

摘要

果蝇中P家族的转座元件被认为是通过一种留下双链缺口的剪切粘贴过程进行转座的。这种缺口的修复导致了多达几千碱基对的信息通过类似于基因转换的过程从同源模板序列转移到P元件切除位点。该模板是一个在体外修饰的序列,在不同的基因组位置进行了测试。对123个转换片段的表征提供了它们长度和分布的详细描述。大多数事件是连续的转换片段,它们与P插入位点重叠,而模板没有伴随转换。平均转换片段为1379个碱基对,片段长度的分布符合缺口扩大的简单模型。转换事件以足够高的频率发生,从而构成了一种有效的定向基因替换方法的基础。

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