DeVault J D, Hughes K J, Leopold R A, Johnson O A, Narang S K
Department of Insect Genetics and Biochemistry, U.S. Department of Agriculture/Agricultural Research Service, Fargo, North Dakota, 58105, USA.
Genome Res. 1996 Jul;6(7):571-9. doi: 10.1101/gr.6.7.571.
Transposable elements with short inverted repeats at their termini have been identified in a number of diverse insect species and have proven to be useful gene delivery vectors for the transformation of Drosophila melanogaster. In this report we examine the ability of the D. melanogaster hobo element to transpose in lepidopteran species. A Trichoplusia ni (cabbage looper) and a Helicoverpa zea (corn earworm) embryonic cell line were found to be capable of supporting productive transposition of the hobo element as measured by a plasmid-based excision assay. Furthermore, hobo transposition was detected in H. zea embryos in a manner consistent with that seen for the cell line. In both cases, transposition/excision was found to be independent of vector-encoded transposase functions, indicating that endogenous genes are involved in hobo mobility. Finally, we demonstrate the stable insertion of the bacterial lacZ gene into the H. zea genome. These data demonstrate that hobo elements are capable of transgressing species boundaries and functioning in non-drosophilid cellular environments. More importantly, this represents the first description of a genetic transformation system for a lepidopteran species.
在许多不同的昆虫物种中都已鉴定出末端带有短反向重复序列的转座元件,并且这些元件已被证明是用于转化黑腹果蝇的有用基因传递载体。在本报告中,我们研究了黑腹果蝇hobo元件在鳞翅目物种中转座的能力。通过基于质粒的切除试验测定,发现粉纹夜蛾(甘蓝夜蛾)和玉米穗虫的胚胎细胞系能够支持hobo元件的有效转座。此外,在玉米穗虫胚胎中检测到hobo转座,其方式与细胞系中所见一致。在这两种情况下,转座/切除均被发现独立于载体编码的转座酶功能,这表明内源基因参与了hobo的移动性。最后,我们证明了细菌lacZ基因稳定插入玉米穗虫基因组。这些数据表明hobo元件能够跨越物种界限并在非果蝇细胞环境中发挥作用。更重要的是,这代表了鳞翅目物种遗传转化系统的首次描述。