Molecular cloning and expression of the mouse ornithine decarboxylase gene.
Molecular cloning and expression of the mouse ornithine decarboxylase gene.
复制标题
小鼠鸟氨酸脱羧酶基因的分子克隆和表达。
DOI：
10.1073/pnas.81.2.540
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发表时间：
1984
影响因子：
11.1
通讯作者：
Coffino,P
中科院分区：
文献类型：
--
作者：
McConlogue,L;Gupta,M;Wu,L;Coffino,P
我们利用以鸟氨酸脱羧酶（OrnDCase）为主要蛋白产物的突变型S49小鼠淋巴瘤细胞系mRNA合成并克隆cDNA。通过OrnDCase mRNA杂交选择和体外翻译鉴定含有OrnDCase cDNA的质粒。插入量最大的两个片段共跨越2.05千碱基的cDNA。用EcoRI或BamHI切割野生型或突变型S49细胞的DNA进行Southern blot分析，发现多个与OrnD-Case cDNA同源的条带，在突变细胞中仅扩增出一条。RNA转移印迹分析显示，小鼠淋巴瘤细胞中主要的OrnD-Case mRNA长度为2.0千碱基。在小鼠肾脏中发现了类似大小的mRNA，并且在睾酮（OrnDCase活性的诱导剂）处理的小鼠肾脏中更为丰富。
We used mRNA from a mutant S49 mouse lymphoma cell line that produces ornithine decarboxylase (OrnDCase) as its major protein product to synthesize and clone cDNA. Plasmids containing OrnDCase cDNA were identified by hybrid selection of OrnDCase mRNA and in vitro translation. The two of these with the largest inserts together span 2.05 kilobases of cDNA. Southern blot analysis of DNA from wild-type or mutant S49 cells, cleaved with EcoRI or with BamHI, revealed multiple bands homologous to OrnD-Case cDNA, only one of which was amplified in the mutant cells. RNA transfer blot analysis showed that the major OrnD-Case mRNA in the mouse lymphoma cells is 2.0 kilobases long. A similar size mRNA was found in mouse kidney and was more abundant in the kidneys of mice treated with testosterone, an inducer of OrnDCase activity in that tissue.