Capone J P, Sharp P A, RajBhandary U L
EMBO J. 1985 Jan;4(1):213-21. doi: 10.1002/j.1460-2075.1985.tb02338.x.
Amber, ochre and opal suppressor tRNA genes have been generated by using oligonucleotide directed site-specific mutagenesis to change one or two nucleotides in a human serine tRNA gene. The amber and ochre suppressor (Su+) tRNA genes are efficiently expressed in CV-1 cells when introduced as part of a SV40 recombinant. The expressed amber and ochre Su+ tRNAs are functional as suppressors as demonstrated by readthrough of the amber codon which terminates the NS1 gene of an influenza virus or the ochre codon which terminates the hexon gene of adenovirus, respectively. Interestingly, several attempts to obtain the equivalent virus stock of an SV40 recombinant containing the opal suppressor tRNA gene yielded virus lacking the opal suppressor tRNA gene. This suggests that expression of an efficient opal suppressor derived from a human serine tRNA gene is highly detrimental to either cellular or viral processes.
通过使用寡核苷酸定向位点特异性诱变来改变人类丝氨酸tRNA基因中的一个或两个核苷酸,已产生了琥珀色、赭石色和乳白抑制tRNA基因。当作为SV40重组体的一部分导入时,琥珀色和赭石色抑制(Su+)tRNA基因在CV-1细胞中高效表达。表达的琥珀色和赭石色Su+ tRNA具有抑制功能,分别通过流感病毒NS1基因终止的琥珀密码子或腺病毒六邻体基因终止的赭石密码子的通读来证明。有趣的是,几次尝试获得含有乳白抑制tRNA基因的SV40重组体的等效病毒株,结果得到的病毒缺乏乳白抑制tRNA基因。这表明源自人类丝氨酸tRNA基因的高效乳白抑制子的表达对细胞或病毒过程具有高度有害性。