In vivo electroporation of DNA into the wing epidermis of the butterfly, Bicyclus anynana.
In vivo electroporation of DNA into the wing epidermis of the butterfly, Bicyclus anynana.
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DOI：
10.1673/031.007.5301
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发表时间：
2007
期刊：
影响因子：
--
通讯作者：
Monteiro A
中科院分区：
文献类型：
--
作者：
Golden K;Sagi V;Markwarth N;Chen B;Monteiro A
将基因直接转移到分化的昆虫组织中是确定基因功能的有用方法，因为它避免了进行生殖系转化和具有关于组织特异性基因启动子的信息的需要。在此，通过将报告基因电穿孔到蝴蝶Bicyclus anynana（Butler）（鳞翅目：蛱蝶科）的蛹前翅中来实现体内基因递送。含有增强型绿色荧光蛋白（EGFP）编码序列的质粒由果蝇热休克启动子hsp 70驱动，在注射质粒后进行电穿孔和热休克后，在表皮细胞中成功表达。EGFP表达仅限于注射和电穿孔位点附近，但转化细胞的数量从几个到超过5000个细胞不等。优化电穿孔参数以使转化细胞的数量最大化，同时使对成虫翅膀的损伤程度最小化。虽然翅膀组织对某些电参数耐受良好，但插入钨电极造成的物理损伤导致穿刺部位周围的成人翅膀图案频繁中断。虽然这种技术可以用于测试标记基因（如EGFP）在新构建的质粒中的正确表达，但其在测试候选基因在翅型形成中的功能方面的潜在用途是有限的。
The direct transfer of genes into differentiated insect tissues is a useful method of determining gene function because it circumvents the need to perform germ line transformations and of having information on tissue-specific gene promoters. Here in vivo gene delivery is achieved through electroporation of a reporter gene into the pupal forewing of the butterfly Bicyclus anynana (Butler) (Lepidoptera: Nymphalidae). Plasmids containing the coding sequence for enhanced green fluorescent protein (EGFP), driven by the Drosophila heat-shock promoter hsp70, were successfully expressed in epidermal cells after plasmid injection followed by electroporation and heat shock. EGFP expression was restricted to the vicinity of the injection and electroporation site, but the number of transformed cells varied from a few to over 5000 cells. Electroporation parameters were optimized in order to maximize the number of transformed cells while minimizing the extent of damage to the adult wing. While certain electrical parameters were well tolerated by the wing tissue, the physical damage caused by the insertion of the tungsten electrodes led to frequent disruptions of the adult wing pattern around the puncture sites. While this technique can be useful to test the correct expression of marker genes (such as EGFP) in newly build plasmids immediately following their injection, its potential use in testing the function of candidate genes in wing pattern formation is limited.
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影响因子：
64.8
作者：
Brakefield, PM;Gates, J;Carroll, SB
通讯作者：
Carroll, SB
影响因子：
1.5
作者：
Tawk, M;Tuil, D;Paulin, D
通讯作者：
Paulin, D
影响因子：
3.9
作者：
Lee, JM;Takahashi, M;Kusakabe, T
通讯作者：
Kusakabe, T
影响因子：
64.5
作者：
Teleman, AA;Cohen, SM
通讯作者：
Cohen, SM
影响因子：
2.7
作者：
Saito, T;Nakatsuji, N
通讯作者：
Nakatsuji, N