Thompson H A, Burson J M, Lang J A, Gross K W, Sigmund C D
Department of Internal Medicine, University of Iowa College of Medicine, Iowa City 5224, USA.
Endocrinology. 1995 Jul;136(7):3037-45. doi: 10.1210/endo.136.7.7789330.
As4.1 cells are derived from a renin-expressing kidney tumor induced by tissue-specific oncogene-mediated tumorigenesis in transgenic mice. These cells express high levels of renin messenger RNA (mRNA) and synthesize prorenin and renin; they were therefore used as a model to further investigate the molecular biology of renin-producing kidney cells by cloning and characterizing novel mRNAs expressed in these cells. One clone, designated 1.5, was randomly selected from an As4.1 complementary DNA (cDNA) library, and two other cDNA clones, designated 4.9 and 6.9, were obtained by screening the cDNA library using a strategy to identify As4.1 cell-specific mRNAs. Each clone exhibited a highly restricted tissue-specific expression profile, including high level expression in As4.1 cells and low level expression in kidney. No homology was found between the sequence of the partial 1.5 and 4.9 cDNAs and sequences in Genbank. Southern blot analysis revealed that clone 4.9 is encoded by a single copy gene containing at least two separate exons. A homology search of the sequence of clone 6.9 revealed it to encode a cDNA to serum amyloid A protein; consistent with this identification, expression of 6.9 mRNA was highly induced in both kidney and liver after treatment of mice with Escherichia coli lipopolysaccharide.
As4.1细胞源自转基因小鼠中由组织特异性癌基因介导的肿瘤发生所诱导的表达肾素的肾肿瘤。这些细胞表达高水平的肾素信使核糖核酸(mRNA)并合成肾素原和肾素;因此,它们被用作模型,通过克隆和鉴定这些细胞中表达的新型mRNA来进一步研究产生肾素的肾细胞的分子生物学。从As4.1互补DNA(cDNA)文库中随机选择一个名为1.5的克隆,并使用鉴定As4.1细胞特异性mRNA的策略筛选cDNA文库,获得另外两个名为4.9和6.9的cDNA克隆。每个克隆都表现出高度受限的组织特异性表达谱,包括在As4.1细胞中的高水平表达和在肾脏中的低水平表达。在Genbank中,1.5和4.9部分cDNA的序列与其他序列之间未发现同源性。Southern印迹分析表明,克隆4.9由一个包含至少两个独立外显子的单拷贝基因编码。对克隆6.9的序列进行同源性搜索发现,它编码一种血清淀粉样蛋白A的cDNA;与此鉴定结果一致,在用大肠杆菌脂多糖处理小鼠后,6.9 mRNA在肾脏和肝脏中均被高度诱导表达。