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鸡肝脏和输卵管mRNA的复杂性与多样性分析

Analysis of the complexity and diversity of mRNA from chicken liver and oviduct.

作者信息

Axel R, Feigelson P, Schutz G

出版信息

Cell. 1976 Feb;7(2):247-54. doi: 10.1016/0092-8674(76)90024-6.

Abstract

We have analyzed the sequence complexity and diversity of poly(A)-containing mRNA derived from two highly differentiated chicken tissues. Two independent approaches were used in our analyses. The first involves the annealing of cDNA copies of mRNA to a vast excess of the template RNA; the second procedure uses hybridization between highly radioactive single-copy genomic DNA and mRNA. The results obtained using these two experimental approaches are in good accord and reveal the presence of 12,000-15,000 diverse mRNA species in both chicken liver and oviduct. In both cell types, the kinetics of annealing of cDNA to its template mRNA demonstrate discrete frequency classes with most of the different mRNA species present in fewer than 10 copies per cell. 70% of oviduct mRNA, however, consists of about 10 abundant RNA species, which probably are responsible for the synthesis of the egg white proteins. The diversity of mRNA species in chicken liver and oviduct was further studied by heterologous annealing reactions between cDNA or singlecopy genomic DNA and a vast excess of mRNA. These studies demonstrate that 85% of the different mRNA sequences detected are present in both liver and oviduct, and suggest that the vast majority of the information expressed as mRNA is required for the maintenance of cellular functions common to all tissues.

摘要

我们分析了来自两种高度分化的鸡组织的含多聚腺苷酸(poly(A))的mRNA的序列复杂性和多样性。我们的分析采用了两种独立的方法。第一种方法是将mRNA的cDNA拷贝与大量过量的模板RNA进行退火;第二种方法是利用高放射性单拷贝基因组DNA与mRNA之间的杂交。使用这两种实验方法获得的结果高度一致,并揭示了鸡肝脏和输卵管中存在12,000 - 15,000种不同的mRNA种类。在这两种细胞类型中,cDNA与其模板mRNA的退火动力学显示出离散的频率类别,大多数不同的mRNA种类在每个细胞中的拷贝数少于10个。然而,输卵管中70%的mRNA由大约10种丰富的RNA种类组成,这些种类可能负责蛋清蛋白的合成。通过cDNA或单拷贝基因组DNA与大量过量mRNA之间的异源退火反应,进一步研究了鸡肝脏和输卵管中mRNA种类的多样性。这些研究表明,所检测到的85%的不同mRNA序列在肝脏和输卵管中都存在,并表明绝大多数以mRNA形式表达的信息是维持所有组织共有的细胞功能所必需的。

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