小鼠新基因Ayu17-449的克隆与表达分析

TANG Hua , ARAKI Kimi , YAMAMURA Ken-ichi
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引用次数: 1

摘要

我们利用基因捕获载体PU8从小鼠胚胎干细胞中寻找一些在小鼠发育过程中起重要作用的有趣基因。捕获了一个阳性ES菌落Ayu17-449。用5′RACE法获得其部分cDNA。与EST数据库中一个5523 bp的cDNA片段(GI: 20879412)同源。进一步分析Celera小鼠基因数据库中5523 bp的cDNA序列,发现该序列与两个基因重叠。我们根据这两个基因的mrna设计了一系列的DNA引物进行RT-PCR和Northern blotting分析,鉴定出一个编码1920 aa的约9 kb的新RNA(我们命名为Ayu17-449)。这种基因在大脑、肾脏、心脏、肺、肌肉和胃中高度表达。表达的蛋白在其n端含有Granin基序,表明该基因可能参与激素分泌。
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Cloning and Expression Analysis of a Murine Novel Gene, Ayu17-449

We used gene trapping vector PU8 to search some interesting genes which play important roles in mouse development from murine ES cells. One positive ES colony termed Ayu17–449 was trapped. Its partial cDNA was obtained by using 5′ RACE method. It is homologous to a 5523 bp cDNA fragment (GI: 20879412) in EST database. Further analysis of the 5523 bp cDNA sequence in Celera mouse gene database showed that it overlaps two genes. We designed serials of DNA primers according to the mRNAs of these two genes for RT-PCR and Northern blotting analysis, and identified a novel RNA about 9 kb (we named it as Ayu17-449) encoding 1920 aa. This gene is expressed highly in the brain, kidney, heart, lung, muscle and stomach. The expressed protein contains a Granin motif on its N-terminus, showing that this gene may be involved in hormone secretion.

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