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酵母中多聚腺苷酸化RNA序列的数量与分布

Number and distribution of polyadenylated RNA sequences in yeast.

作者信息

Hereford L M, Rosbash M

出版信息

Cell. 1977 Mar;10(3):453-62. doi: 10.1016/0092-8674(77)90032-0.

Abstract

The poly(A)-containing RNA, isolated from the budding yeast Saccharomyces cerevisiae, has been characterized with regard to the number and distribution of sequences by a kinetic analysis of RNA-cDNA hybridization. In agreement with results previously obtained on metazoan eucaryotes (Bishop et al., 1974), discrete complexity classes were observed. There exist low, medium, and high complexity classes which contain approximately 20, 400, and 2400 sequences, respectively. This measurements of the number of sequences has been verified by hybridization with single copy DNA. 20% of the single copy fraction of the yeast genome is rendered double-stranded by poly(A)-containing RNA. Assuming asymmetric transcription, this is equivalent to approximately 4000 poly(A)-containing sequences, verifying the results obtained with RNA-cDNA hybridization. In addition, the first-order kinetics of the hybridization with single copy DNA verified the notion that most of the sequence complexity is present at the same intracellular concentration. The same number and distribution of sequences were found in poly(A)-containing polysomal RNA and in total RNA, suggesting that most or all of the sequence complexity is on polysomes and is adenylated. The results indicate that RNA-cDNA hybridization is an accurate method for determining sequence complexity values and that yeast, grown under vegetative conditions, has 3000-4000 different mRNA sequences.

摘要

从出芽酵母酿酒酵母中分离得到的含聚腺苷酸(poly(A))的RNA,已通过RNA - cDNA杂交的动力学分析,在序列数量和分布方面进行了表征。与先前在后生真核生物上获得的结果一致(Bishop等人,1974年),观察到了离散的复杂性类别。存在低、中、高复杂性类别,分别包含约20、400和2400个序列。通过与单拷贝DNA杂交,对序列数量的测量得到了验证。酵母基因组单拷贝部分的20%被含poly(A)的RNA双链化。假设不对称转录,这相当于约4000个含poly(A)的序列,验证了RNA - cDNA杂交获得的结果。此外,与单拷贝DNA杂交的一级动力学验证了这样一种观点,即大多数序列复杂性以相同的细胞内浓度存在。在含poly(A)的多聚核糖体RNA和总RNA中发现了相同数量和分布的序列,这表明大多数或所有序列复杂性都存在于多聚核糖体上并且被腺苷酸化。结果表明,RNA - cDNA杂交是确定序列复杂性值的准确方法,并且在营养条件下生长的酵母具有3000 - 4000种不同的mRNA序列。

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