The piggyBac element is capable of precise excision and transposition in cells and embryos of the mosquito, Anopheles gambiae

被引:31
|
作者
Grossman, GL
Rafferty, CS
Fraser, MJ
Benedict, MQ
机构
[1] Ctr Dis Control & Prevent, Div Parasit Dis, Entomol Branch, Atlanta, GA 30341 USA
[2] Univ Notre Dame, Dept Biol, Notre Dame, IN 46556 USA
关键词
Anopheles gambiae; piggyBac; Mos55; transposable elements; transposition assay; transformation; mosquitoes;
D O I
10.1016/S0965-1748(00)00092-8
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The piggyBac transposable element was tested for transposition activity in plasmid-based excision and inter-plasmid transposition assays to determine if this element would function in Anopheles gambiae cells and embryos. In the Mos55 cell line, precise excision of the piggyBac element was observed only in the presence of a helper plasmid. Excision occurred at a rate of 1 event per 1000 donor plasmids screened. Precise excision of the piggyBac element was also observed in injected An. gambiae embryos, but at a lower rate of 1 excision per 5000 donor plasmids. Transposition of the marked piggyBac element into a target plasmid occurred in An. gambiae cells at a rate of 1 transposition event per 24,000 donor plasmids. The piggyBac element transposed in a precise manner, with the TTAA target site being duplicated upon insertion, in 56% of transpositions observed, and only in the presence of the piggyBac helper. The remaining transpositions resulted in a deletion of target sequence, a novel observation for the phenomenon of piggyBac element insertion. 'Hot spots' for insertion into the target plasmid were observed, with 25 of 34 events involving one particular site. These results are the first demonstration of the precise mobility of piggyBac in this malaria vector and suggest that the lepidopteran piggyBac transposon is a candidate element for germline transformation of anopheline mosquitoes. Published by Elsevier Science Ltd.
引用
收藏
页码:909 / 914
页数:6
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