The lepidopteran transposon vector, piggyBac, mediates germ-line transformation in the Mediterranean fruit fly
The lepidopteran transposon vector, piggyBac, mediates germ-line transformation in the Mediterranean fruit fly
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DOI：
10.1073/pnas.95.13.7520
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发表时间：
1998-06-23
影响因子：
11.1
通讯作者：
Saul, SH
中科院分区：
文献类型：
--
作者：
Handler, AM;McCombs, SD;Saul, SH
在地中海实蝇Ceratitis capitata中测试来自甘蓝夜蛾粉纹夜蛾的piggyBac（IFP 2）短反向末端重复转座元件作为二分载体辅助系统的一部分的基因转移载体功能。在白色眼宿主菌株中，用两种不同浓度的正常调节的piggyBac转座酶辅助物测试用地中海实蝇白色基因标记的piggyBac载体。两个实验都以大约3- 5%的频率产生转化体，总共分离出6个具有不同着色水平的有色眼睛的品系，来自每个品系的G（1）染色体同胞具有至少一个共同的整合，几个亚系具有额外的第二整合。对于分离的第一个转基因株系，两个整合被确定为稳定15代。对于五个系，通过对通过反向PCR分离的插入位点连接进行测序来验证piggyBac介导的转座，所述反向PCR鉴定了特征性piggyBac TTAA靶位点重复。用鳞翅目载体对地中海果蝇的有效和稳定的转化代表了在相对大的进化距离上的转座子功能，并表明piggyBac系统将在广泛的昆虫中起作用。
The piggyBac (IFP2) short inverted terminal repeat transposable element from the cabbage looper Trichoplusia ni was tested for gene transfer vector function as part of a bipartite vector-helper system in the Mediterranean fruit fly Ceratitis capitata, A piggyBac vector marked with the medfly white gene was tested with a normally regulated piggyBac transposase helper at two different concentrations in a white eye host strain. Both experiments yielded transformants at an approximate frequency of 3-5%, with a total of six lines isolated having pigmented eyes with various levels of coloration, G(1) transformant siblings from each line shared at least one common integration, with several sublines having an additional second integration. For the first transformant line isolated, two integrations were determined to be stable for 15 generations. For five of the lines, a piggyBac-mediated transposition,vas verified by sequencing the insertion site junctions isolated by inverse PCR that identified a characteristic piggyBac TTAA target site duplication. The efficient and stable transformation of the medfly with a lepidopteran vector represents transposon function over a relatively large evolutionary distance and suggests that the piggyBac system will be functional in a broad range of insects.