Young J F, Capecchi M, Laski F A, RajBhandary U L, Sharp P A, Palese P
Science. 1983 Aug 26;221(4613):873-5. doi: 10.1126/science.6308765.
Transfer RNA (tRNA) suppression of nonsense mutations in prokaryotic systems has been widely used to study the structure and function of different prokaryotic genes. Through genetic engineering techniques, it is now possible to introduce suppressor (Su+) tRNA molecules into mammalian cells. A quantitative assay of the suppressor tRNA activity in these mammalian cells is described; it is based on the amount of tRNA-mediated readthrough of a terminating codon in the influenza virus NS1 gene after the cells are infected with virus. Suppressor activity in L cells continuously expressing Su+ (tRNAtyr) was 3.5 percent and that in CV-1 cells infected with an SV40- Su+ (tRNAtyr) recombinant was 22.5 percent.
转运RNA(tRNA)对原核系统中无义突变的抑制作用已被广泛用于研究不同原核基因的结构和功能。通过基因工程技术,现在已能够将抑制性(Su+)tRNA分子导入哺乳动物细胞。本文描述了一种对这些哺乳动物细胞中抑制性tRNA活性的定量测定方法;该方法基于细胞感染病毒后,tRNA介导的流感病毒NS1基因终止密码子通读的量。持续表达Su+(tRNAtyr)的L细胞中的抑制活性为3.5%,而感染SV40 - Su+(tRNAtyr)重组体的CV - 1细胞中的抑制活性为22.5%。