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The piggyBac transposon mediates germ-line transformation in the Orientalfruit fly and closely related elements exist in its genome

机译:piggyBac转座子介导了东方果蝇的种系转化,并且其基因组中存在密切相关的元素

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Germ-line transformation of a white eye strain of the Oriental fruit fly, Bactrocera dorsalis, was achieved with the piggyBac vector, derived from a transposon originally isolated from the cabbage looper moth, Trichoplusia ni. The Vector was marked with the medfly white(+) gene cDNA, and three transgenic lines were identified at a frequency of approximately 2% per fertile G(0). Vector integrations were verified by Southern DNA hybridization, which also revealed the presence of endogenous genomic elements closely related to piggyBac. Approximately 10-20 elements per genome were evident in several B. dorsalis strains, and sequence analysis of 1.5 kb gene amplification products from two wild strains and the white eye host strain indicated 95% nucleotide and 92% amino acid sequence identity among resident elements and the T. ni element. PiggyBac was not evident by hybridization in other tephritid species, or insects previously transformed with the transposon, This is the first discovery of piggyBac beyond T. ni, and its existence in a distantly related species has important implications for the practical use of the vector and insects transformed with it.
机译:使用piggyBac载体实现了东方实蝇白实蝇Bactrocera dorsalis的白眼菌株的种系转化,该载体衍生自最初从白菜loop蛾Trichoplusia ni中分离出的转座子。用medfly white(+)基因cDNA标记载体,并以每个可育G(0)约2%的频率鉴定了三个转基因品系。载体整合已通过Southern DNA杂交进行了验证,该DNA杂交还揭示了与piggyBac密切相关的内源性基因组元件的存在。在几个背侧双歧杆菌菌株中每个基因组大约有10-20个元素,并且对来自两个野生菌株和白眼宿主菌株的1.5 kb基因扩增产物进行的序列分析表明,常驻元件之间存在95%的核苷酸和92%的氨基酸序列同一性。 T. ni元素。 PiggyBac不能通过与其他拟除虫菊属物种的杂交或以前用转座子转化的昆虫来发现。这是除T. ni以外的第一个piggyBac发现,它在远缘相关物种中的存在对载体的实际使用和昆虫随它而变。

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