Germline transformation of Drosophila virilis with the transposable element mariner.
Germline transformation of Drosophila virilis with the transposable element mariner.
复制标题
用转座元件 mariner 进行果蝇 virilis 的种系转化。
DOI：
10.1093/genetics/143.1.365
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发表时间：
1996
期刊：
影响因子：
3.3
通讯作者：
Hartl,DL
中科院分区：
文献类型：
--
作者：
Lohe,AR;Hartl,DL
分子遗传学的一个重要目标是鉴定可作为多种昆虫的有效转化载体的转座元件。转座因子mariner天然存在于多种昆虫中。虽然实际上所有mariner元件都是无功能的，但从毛里求斯果蝇中分离出的Mosl元件是有功能的。Mosl被注射到果蝇胚胎的极细胞区域。 virilis，最后与D.有共同的祖先。毛里求斯四千万年前。在注射动物后代的几个 DNA 库中检测到 MoslPCR 片段，并建立了回交系。因为 Golines 是汇集在一起的，所以实际上可能只获得了一个转化事件，产生的最小频率为 4%。Mosl 以孟德尔方式分离，证明了染色体整合。拷贝数通过自发动员而增加。原位杂交证实了Mosl的多个多态性位置。整合导致特征性的2-bp TA重复。一个 Mosl 元素整合到 370 bp 重复序列的串联阵列中。一些拷贝可能已经整合到异染色质中，正如它们支持 PCR 扩增的能力所证明的那样，尽管在 Southern 和原位杂交中没有信号。
An important goal in molecular genetics has been to identify a transposable element that might serve as an efficient transformation vector in diverse species of insects. The transposable elementmarineroccurs naturally in a wide variety of insects. Although virtually allmarinerelements are nonfunctional, theMoslelement isolated fromDrosophila mauritianais functional.Moslwas injected into the pole-cell region of embryos ofD. virilis, which last shared a common ancestor withD. mauritiana40 million years ago.MoslPCR fragments were detected in several pools of DNA from progeny of injected animals, and backcross lines were established. Because Golines were pooled, possibly only one transformation event was actually obtained, yielding a minimum frequency of 4%.Moslsegregated in a Mendelian fashion, demonstrating chromosomal integration. The copy number increased by spontaneous mobilization.In situhybridization confirmed multiple polymorphic locations ofMosl.Integration results in a characteristic 2-bp TA duplication. OneMoslelement integrated into a tandem array of 370-bp repeats. Some copies may have integrated into heterochromatin, as evidenced by their ability to support PCR amplification despite absence of a signal in Southern andin situhybridizations.