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果蝇多线DNA和RNA中聚腺苷酸序列的大小及分布

Size and distribution of polyadenylic acid sequences in Drosophila polytene DNA and RNA.

作者信息

Alonso C, Pages M, García M L

出版信息

Biochim Biophys Acta. 1977 Dec 2;479(3):235-45. doi: 10.1016/0005-2787(77)90106-x.

Abstract

[3H]Poly(U) hybridizes very rapidly to polytene DNA from Drosophila hydei. When hybridization is performed at 30 degrees C in 2 X SSC to a large excess of DNA, 95% of the poly(U) becomes ribonuclease resistant. Also, complementary RNA transcribed in vitro from polytene DNA hybridizes to poly(U). 023--0.25% of the DNA is composed of (dA)-rich sequences and 0.23--0.31% of cRNA hybridizes to [3H]poly(U). The length of the (dA)-rich sequences on the DNA and cRNA is 40 nucleotides. The Tm values of these hybrids formed between DNA or cRNA-poly(U) is 45 degrees C. The poly(A) fragments from cytoplasmic RNA ranged from 80 to 170 nucleotides in lenght, and migrated in polyacrilamide gels as a broad peak. The average sizes of the poly(A) fragments from the poly(A)-containing RNA transcribed by nuclei isolated from salivary glands in vivo or in vitro were 40, 70, 170 and 70 nucleotides, respectively. Hybridization in situ of [3H]-poly(U) to chromosome squashes indicated that the (dA)-rich sequences are randomly distributed over the whole genome.

摘要

[3H]聚尿苷酸(Poly(U))能非常迅速地与海德氏果蝇(Drosophila hydei)的多线DNA杂交。当在30℃下于2×SSC中对大量过量的DNA进行杂交时,95%的聚尿苷酸变得对核糖核酸酶有抗性。此外,从多线DNA体外转录的互补RNA也能与聚尿苷酸杂交。0.23% - 0.25%的DNA由富含(dA)的序列组成,且0.23% - 0.31%的互补RNA能与[3H]聚尿苷酸杂交。DNA和互补RNA上富含(dA)序列的长度为40个核苷酸。DNA或互补RNA - 聚尿苷酸之间形成的这些杂交体的解链温度(Tm)值为45℃。细胞质RNA中的聚腺苷酸(poly(A))片段长度在80至170个核苷酸之间,在聚丙烯酰胺凝胶中迁移形成一个宽峰。体内或体外从唾液腺分离的细胞核转录的含聚腺苷酸RNA中的聚腺苷酸片段的平均大小分别为40、70、170和70个核苷酸。[3H]聚尿苷酸与染色体压片的原位杂交表明富含(dA)的序列随机分布于整个基因组。

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