转座因子
生物
黑腹果蝇
重组酶
遗传学
整合酶
基因
P元素
黑腹菌
计算生物学
表情盒
基因组
转座酶
重组DNA
载体(分子生物学)
重组
作者
Koen J. T. Venken,Karen L. Schulze,Nele A. Haelterman,Hongling Pan,Yuchun He,Martha Evans-Holm,Joseph W. Carlson,Robert Levis,Allan C. Spradling,Roger A. Hoskins,Hugo J. Bellen
出处
期刊:Nature Methods
[Springer Nature]
日期:2011-07-31
卷期号:8 (9): 737-743
被引量:694
摘要
Described is a randomly inserting transposon that can be swapped for gene traps, coding insertions or protein tag genes, thus expanding the toolkit for Drosophila melanogaster genome engineering. We demonstrate the versatility of a collection of insertions of the transposon Minos-mediated integration cassette (MiMIC), in Drosophila melanogaster. MiMIC contains a gene-trap cassette and the yellow+ marker flanked by two inverted bacteriophage ΦC31 integrase attP sites. MiMIC integrates almost at random in the genome to create sites for DNA manipulation. The attP sites allow the replacement of the intervening sequence of the transposon with any other sequence through recombinase-mediated cassette exchange (RMCE). We can revert insertions that function as gene traps and cause mutant phenotypes to revert to wild type by RMCE and modify insertions to control GAL4 or QF overexpression systems or perform lineage analysis using the Flp recombinase system. Insertions in coding introns can be exchanged with protein-tag cassettes to create fusion proteins to follow protein expression and perform biochemical experiments. The applications of MiMIC vastly extend the D. melanogaster toolkit.
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