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导入小鼠成肌细胞和非成肌细胞的鸡肌动蛋白基因的表达与调控

Expression and regulation of chicken actin genes introduced into mouse myogenic and nonmyogenic cells.

作者信息

Seiler-Tuyns A, Eldridge J D, Paterson B M

出版信息

Proc Natl Acad Sci U S A. 1984 May;81(10):2980-4. doi: 10.1073/pnas.81.10.2980.

Abstract

We have introduced the chicken genes for cytoplasmic beta-actin, cardiac alpha-actin, and skeletal alpha-actin into C2 cells, a murine myogenic cell line, and into L cells by using the simian virus 40-derived vector PSV2 -gpt. In each selection, the entire population of transformed cells was analyzed for the expression and regulation of the actin genes by nuclease S1 assay and primer extension. This was compared to the expression of the vector marker Eco-gpt. The beta-actin gene is transcribed accurately and efficiently both in L-cells and in undifferentiated C2 cells. In fused C2 cells, beta-actin transcripts decrease significantly in parallel with the endogenous level of mouse beta-actin mRNA. Eco-gpt RNA levels remain essentially constant during myogenesis. The alpha-actin genes are correctly expressed at low levels in L cells but at significantly higher levels in the C2 cell background. Unlike the endogenous mouse alpha-actin gene, this level of expression does not change measurably with myogenesis. The skeletal alpha-actin gene is expressed poorly in pre- and post-fusion C2 cells, displaying no induction with differentiation. These results suggest that the tissue specificity of expression is maintained but the pattern of gene regulation for the sarcomeric actins is not. Factors in addition to the sequences flanking these genes are important for modulating gene expression during development. The decrease in the levels of beta-actin RNA during C2 cell differentiation provides a model system in which to study gene repression during development.

摘要

我们利用猿猴病毒40衍生载体PSV2 -gpt,将鸡的细胞质β-肌动蛋白、心肌α-肌动蛋白和骨骼肌α-肌动蛋白基因导入小鼠成肌细胞系C2细胞以及L细胞中。在每次筛选中,通过核酸酶S1分析和引物延伸法,对整个转化细胞群体的肌动蛋白基因表达和调控情况进行分析,并与载体标记物Eco - gpt的表达进行比较。β-肌动蛋白基因在L细胞和未分化的C2细胞中均能准确且高效地转录。在融合的C2细胞中,β-肌动蛋白转录本显著减少,与小鼠β-肌动蛋白mRNA的内源性水平平行下降。Eco - gpt RNA水平在肌生成过程中基本保持恒定。α-肌动蛋白基因在L细胞中低水平正确表达,但在C2细胞背景中表达水平显著更高。与内源性小鼠α-肌动蛋白基因不同,这种表达水平在肌生成过程中没有明显变化。骨骼肌α-肌动蛋白基因在融合前和融合后的C2细胞中表达较差,在分化过程中未表现出诱导作用。这些结果表明,基因表达的组织特异性得以维持,但肌节肌动蛋白的基因调控模式并非如此。除了这些基因侧翼序列之外的其他因素,对于发育过程中基因表达的调节很重要。C2细胞分化过程中β-肌动蛋白RNA水平的下降提供了一个研究发育过程中基因抑制的模型系统。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7de4/345204/464f7a1501a6/pnas00611-0045-a.jpg

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