Raval P, Iversen P L, Bresnick E
Department of Biochemistry, University of Nebraska Medical Center, Omaha.
Biochem Pharmacol. 1991 Jun 1;41(11):1719-23. doi: 10.1016/0006-2952(91)90175-5.
Two oligodeoxyribonucleotides were synthesized that were specific for the messenger RNAs for the polycyclic hydrocarbon-inducible cytochromes P450IA1 and P450IA2. The solution hybridization technique was modified for the use of these oligodeoxyribonucleotide probes so as to increase the sensitivity and specificity of this method. Using this technique, the steady-state levels of the mRNAs for cytochromes P450IA1 and P450IA2 in control rat liver were determined to be less than 3 and 6 molecules/cell, and 1.8 and 4.0 attomol/micrograms poly (A)+ RNA, respectively. At 15 hr after induction with 3-methylcholanthrene, the steady-state levels of the mRNAs for P450IA1 and P450IA2 were 68 and 200 molecules/cell, and 41.6 and 123 attomol/micrograms poly (A)+ RNA.
合成了两种寡脱氧核糖核苷酸,它们对多环烃诱导的细胞色素P450IA1和P450IA2的信使RNA具有特异性。对溶液杂交技术进行了改进,以使用这些寡脱氧核糖核苷酸探针,从而提高该方法的灵敏度和特异性。使用该技术,测定对照大鼠肝脏中细胞色素P450IA1和P450IA2的信使RNA的稳态水平分别小于3和6个分子/细胞,以及1.8和4.0阿托摩尔/微克聚(A)+RNA。在用3-甲基胆蒽诱导15小时后,P450IA1和P450IA2的信使RNA的稳态水平分别为68和200个分子/细胞,以及41.6和123阿托摩尔/微克聚(A)+RNA。