Lazar E, Jacob M, Krol A, Branlant C
Nucleic Acids Res. 1982 Feb 25;10(4):1193-201. doi: 10.1093/nar/10.4.1193.
A DNA fragment containing a 16 nucleotide sequence mimicking the intron extremities of premessenger RNA aligned as proposed previously (1,2) in a model of splicing mechanism was prepared and used as a probe for accessibility of the 5' extremity of U1 RNA. Hybridization of U1 RNA to the probe under non denaturing conditions and digestion of the hybrid with RNase H revealed that the sequence of U1 RNA which is complementary to the extremities of introns is accessible to hybridization and to enzymes. Therefore, the configuration of isolated U1 RNA satisfies the criteria required for the alignment of introns and further enzymatic reactions of splicing.
制备了一个DNA片段,其包含一段16个核苷酸的序列,该序列模拟了前体信使RNA的内含子末端,按照先前在剪接机制模型中提出的方式(1,2)进行排列,并用作U1 RNA 5'末端可及性的探针。在非变性条件下将U1 RNA与该探针杂交,并用RNase H消化杂交体,结果显示与内含子末端互补的U1 RNA序列可用于杂交和酶作用。因此,分离出的U1 RNA的构型满足内含子排列及剪接进一步酶促反应所需的标准。