A visible dominant marker for insect transgenesis.

A visible dominant marker for insect transgenesis.
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DOI:
10.1038/ncomms2312
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发表时间:
2012
影响因子:
16.6
通讯作者:
--
中科院分区:
综合性期刊1区
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--
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目前大多数昆虫的转基因依赖于荧光标记。在这里,我们建立了一个转化标记系统,由于黑色素颜色的变化,这是广泛存在于动物中,导致肉眼可见的表型。在家蚕中普遍过量表达芳基烷基胺-N-乙酰基转移酶(arylalkylamine-N-acetyltransferase),将新孵化的一龄幼虫的颜色从黑色改变为独特的浅棕色,并且可以通过直接连接到杆状病毒立即早期1基因启动子而用作分子标记。通过BM-芳基烷基胺-N-乙酰基转移酶可以在整个幼虫阶段和成年动物中观察到黑色色素沉着的抑制。或者,在另一个基因B中过表达。mori β-丙氨酰-多巴胺合成酶(Bm-ebony)改变老龄期幼虫的体色,尽管一龄幼虫具有正常的深色。我们进一步表明,异位BM-芳基烷基胺-N-乙酰基转移酶的表达减轻着色瓢虫异色瓢虫和果蝇黑腹果蝇,突出了潜在的有用性,这种标记在不同的昆虫类群的转基因。 昆虫的遗传修饰主要涉及使用荧光标记来识别成功的转化。Osanai-Futahashi等人报道了一种基于黑色素色素变化的标记系统,该系统允许用肉眼识别转基因昆虫。
Transgenesis of most insects currently relies on fluorescence markers. Here we establish a transformation marker system causing phenotypes visible to the naked eye due to changes in the color of melanin pigments, which are widespread in animals. Ubiquitous overexpression of arylalkylamine-N-acetyl transferase in the silkworm, Bombyx mori, changes the color of newly hatched first-instar larvae from black to a distinctive light brown color, and can be used as a molecular marker by directly connecting to baculovirus immediate early 1 gene promoter. Suppression of black pigmentation by Bm-arylalkylamine-N-acetyl transferase can be observed throughout the larval stages and in adult animals. Alternatively, overexpression in another gene, B. mori β-alanyl-dopamine synthetase (Bm-ebony), changes the larval body color of older instars, although first-instar larvae had normal dark coloration. We further show that ectopic Bm-arylalkylamine-N-acetyl transferase expression lightens coloration in ladybird beetle Harmonia axyridis and fruit fly Drosophila melanogaster, highlighting the potential usefulness of this marker for transgenesis in diverse insect taxa. Genetic modification in insects mostly involves the use of fluorescent markers to identify successful transformation. Here Osanai-Futahashi et al. report a marker system based on changes in melanin pigmentation that allows the identification of genetically modified insects with the naked eye.
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发表时间: 1998-07-01
影响因子: 3.1
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