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一种基于双顺反子信使翻译的细胞自主性测试。

A test for cell autonomy, based on di-cistronic messenger translation.

作者信息

Hart K, Bienz M

机构信息

MRC Laboratory of Molecular Biology, Cambridge, UK.

出版信息

Development. 1996 Mar;122(3):747-51. doi: 10.1242/dev.122.3.747.

Abstract

We have devised a test for cell autonomy of a gene that is switched on ectopically in a clone of cells, allowing us to ask whether the wild-type activity of this gene can influence neighbouring cells. To switch on the test gene, we used the yeast FRT system, and marked the FRT-generated cell clone by co-expressing beta-galactosidase. Co-expression is achieved by a stretch of 5' untranslated mRNA from the homeotic gene Ultrabithorax (Ubx), which is inserted between the two coding sequences. We show that this Ubx sequence mediates efficient and reliable di-cistronic mRNA translation in wing imaginal discs of Drosophila. Applying our test to Ubx, we find that ectopic Ubx in wing discs strictly coincides with beta-galactosidase expression. Consequently, wing cells are transformed into cells that appear to be intermediates between wing and haltere cells, contesting the view that homeotic genes act as binary switches.

摘要

我们设计了一种针对在细胞克隆中异位激活的基因的细胞自主性测试,借此可以探究该基因的野生型活性是否能影响邻近细胞。为激活测试基因,我们使用了酵母FRT系统,并通过共表达β-半乳糖苷酶来标记由FRT产生的细胞克隆。共表达是通过一段来自同源异型基因超双胸(Ubx)的5'非翻译mRNA实现的,该序列插入在两个编码序列之间。我们发现,这段Ubx序列在果蝇翅成虫盘中介导了高效且可靠的双顺反子mRNA翻译。将我们的测试应用于Ubx,我们发现在翅盘中异位表达的Ubx与β-半乳糖苷酶的表达严格一致。因此,翅细胞转变为了似乎是翅细胞和平衡棒细胞之间中间态的细胞,这对同源异型基因作为二元开关的观点提出了质疑。

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