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烟草植株叶片中细胞核和多核糖体RNA的序列复杂性

Sequence complexity of nuclear and polysomal RNA in leaves of the tobacco plant.

作者信息

Goldberg R B, Hoschek G, Kamalay J C, Timberlake W E

出版信息

Cell. 1978 May;14(1):123-31. doi: 10.1016/0092-8674(78)90307-0.

Abstract

The first measurements are reported of the sequence complexity of nuclear and polysomal RNA contained within the cells of a higher plant. Polysomal RNA from tobacco leaves was prepared by a procedure which minimized contamination with nuclear RNA. Hybridization of 3H-cDNA complementary to polysomal poly(A) RNA with an excess of tobacco DNA indicated that greater than 95% of the poly(A) mRNA was transcribed from single-copy sequences. RNA excess hybridization reactions with polysomal poly(A) RNA and 3H-cDNA revealed the presence of three abundance classes in the poly(A) mRNA. The best least-squares solution indicated that these classes comprise 9, 52, and 39% of the poly (A) mRNA and contain sequences present an average of 4500, 340 and 17 times per cell. Hybridization reactions containing an excess of nuclear or total polysomal RNA and 3H-single-copy DNA indicated the complexity of these RNA populations to be 1.19 X 10(8) nucleotides (nuclear) and 3.33 X 10(7) nucleotides (polysomal). Thus only 28% of the nuclear RNA sequence diversity (27,000 average-sized mRNA sequences) is respresented in leaf polysomes. These results suggest that there is a general similarity in the basic transcriptional processes of metaphytan and metazoan cells.

摘要

首次报道了高等植物细胞内核糖体RNA和多核糖体RNA序列复杂性的测量结果。烟草叶片的多核糖体RNA是通过一种将核RNA污染降至最低的方法制备的。与多核糖体多聚腺苷酸RNA互补的3H-cDNA与过量烟草DNA的杂交表明,超过95%的多聚腺苷酸mRNA是从单拷贝序列转录而来的。多核糖体多聚腺苷酸RNA与3H-cDNA的RNA过量杂交反应揭示了多聚腺苷酸mRNA中存在三个丰度等级。最佳最小二乘解表明,这些等级分别占多聚腺苷酸mRNA的9%、52%和39%,其序列在每个细胞中平均出现4500次、340次和17次。含有过量核RNA或总多核糖体RNA以及3H单拷贝DNA的杂交反应表明,这些RNA群体的复杂性分别为1.19×10⁸个核苷酸(核RNA)和3.33×10⁷个核苷酸(多核糖体RNA)。因此,叶多核糖体中仅代表了核RNA序列多样性的28%(平均大小的27000个mRNA序列)。这些结果表明,后生植物和后生动物细胞的基本转录过程存在普遍相似性。

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