Savage M J, Sala-Trepat J M, Bonner J
Biochemistry. 1978 Feb 7;17(3):462-7. doi: 10.1021/bi00596a014.
The complexity of rat liver poly (A)+ messenger RNA (mRNA) has been measured by analysis of the kinetics of hydridization with both complementary DNA (cDNA) and single copy DNA. The complementary DNA-poly(A)+ mRNA hybridization reaction demonstrates the existence of three abundance classes representing 18, 37, and 45% of the cDNA and 4, 290, and 24 000 different 1800-nucleotide sequences respectively. The poly(A)+ mRNA driven single copy DNA hybridization reaction reveals a single major transition accounting for 1.9% of the haploid rat genome. The kinetics of the poly(A)+ mRNA driven single copy DNA reaction suggest that approximately 45% of the mass of the mRNA population contains over 95% of the complexity. Although higher than previous estimates, the base sequence complexities of rat liver poly(A)+ mRNA measured in these two ways are in good agreement, suggesting that the technique of poly(A)+ mRNA-cDNA hybridization may be used in approximating the complexity as well as abundance of a messenger RNA population. DNA-driven cDNA reactions reveal that about 10% of rat liver poly(A)+ mRNA is transcribed from repetitive sequences in the rat genome.
通过分析与互补DNA(cDNA)和单拷贝DNA杂交的动力学,对大鼠肝脏多聚腺苷酸加尾信使核糖核酸(mRNA)的复杂性进行了测定。互补DNA与多聚腺苷酸加尾mRNA的杂交反应表明存在三个丰度类别,分别占cDNA的18%、37%和45%,以及分别代表4种、290种和24000种不同的1800个核苷酸序列。多聚腺苷酸加尾mRNA驱动的单拷贝DNA杂交反应显示出一个主要转变,占单倍体大鼠基因组的1.9%。多聚腺苷酸加尾mRNA驱动的单拷贝DNA反应动力学表明,大约45%的mRNA群体质量包含超过95%的复杂性。尽管高于先前的估计,但通过这两种方法测得的大鼠肝脏多聚腺苷酸加尾mRNA的碱基序列复杂性吻合良好,这表明多聚腺苷酸加尾mRNA - cDNA杂交技术可用于近似信使RNA群体的复杂性和丰度。DNA驱动的cDNA反应表明,大鼠肝脏多聚腺苷酸加尾mRNA约10%是由大鼠基因组中的重复序列转录而来。