Schetelig Marc F, Handler Alfred M
U.S. Department of Agriculture, Agricultural Research Service, Center for Medical, Agricultural and Veterinary Entomology, Gainesville, Florida 32608
U.S. Department of Agriculture, Agricultural Research Service, Center for Medical, Agricultural and Veterinary Entomology, Gainesville, Florida 32608.
G3 (Bethesda). 2013 Apr 9;3(4):687-693. doi: 10.1534/g3.112.005488.
Transposable elements are widely used as vectors for integrating transgenes into the genome of insects. However, the random nature of transposon vector integrations often results in mutations and makes transgene expression subject to variable genomic position effects. This makes reliable quantitative comparisons of different transgenes difficult and development of highly fit transgenic strains laborious. Tools for site-specific transgene targeting are essential for functional genomic comparisons and to develop the most advanced transgenic insect strains for applied use. Here we describe a recombinase-mediated cassette exchange gene targeting system based on Cre/loxP that is highly efficient in Drosophila, and for the first time in a non-drosophilid, the tephritid fly, Anastrepha suspensa This system allowed a comparison of the Drosophila constitutive polyubiquitin promoter and the artificial 3xP3 tissue-specific promoter in the same genomic context within each species, showing that the widely used 3xP3 promoter is apparently nonfunctional in the tephritid fly.
转座元件被广泛用作将转基因整合到昆虫基因组中的载体。然而,转座子载体整合的随机性常常导致突变,并使转基因表达受到可变的基因组位置效应的影响。这使得不同转基因之间进行可靠的定量比较变得困难,并且培育高度适应的转基因品系也很费力。用于位点特异性转基因靶向的工具对于功能基因组比较以及开发用于实际应用的最先进转基因昆虫品系至关重要。在此,我们描述了一种基于Cre/loxP的重组酶介导的盒式交换基因靶向系统,该系统在果蝇中效率很高,并且在非果蝇类实蝇Anastrepha suspensa中首次实现。该系统允许在每个物种的相同基因组背景下比较果蝇组成型多聚泛素启动子和人工3xP3组织特异性启动子,结果表明广泛使用的3xP3启动子在实蝇中显然无功能。