Taguchi F, Ikeda T, Makino S, Yoshikura H
National Institute of Neuroscience, NCNP, Tokyo, Japan.
Virology. 1994 Jan;198(1):355-9. doi: 10.1006/viro.1994.1041.
A plaque-cloned mouse hepatitis virus mutant, MHV-S No. 8, was isolated from Ki-BALB cells persistently infected with MHV-S. The mRNAs 1 to 6 were larger in the mutant, whereas there was no difference between the two viruses in the size of the smallest mRNA, mRNA 7. Sequence analyses of the genomic RNA, mRNA 6, and mRNA 7 of the two viruses revealed that an additional 111 nt were inserted just upstream of the intergenic consensus sequence preceding the N gene in MHV-S No. 8. The inserted region consisted of two different parts; the 3'-most 30 nt corresponded to nucleotides 28 to 57 of the leader sequence and the 5'-most 81 nt corresponded to nucleotides 58 to 138 of mRNA 7. This structure of No. 8 was most likely generated by RNA-RNA recombination between genomic RNA and subgenomic RNA species. The nucleotide insertion in the intergenic sequence between genes M and N resulted in two consensus sequences separated by 111 nt. Primer extension analysis revealed that the amount of a slightly larger, subgenomic mRNA resulting from initiation of synthesis at the upstream consensus sequence was only 5% of the usual sized mRNA 7 initiated from the downstream consensus sequence.
从持续感染MHV-S的Ki-BALB细胞中分离出一种噬菌斑克隆的小鼠肝炎病毒突变体MHV-S No. 8。突变体中的mRNA 1至6更大,而两种病毒中最小的mRNA即mRNA 7的大小没有差异。对两种病毒的基因组RNA、mRNA 6和mRNA 7进行序列分析发现,在MHV-S No. 8的N基因之前的基因间共有序列上游刚好插入了额外的111个核苷酸。插入区域由两个不同部分组成;最3'端的30个核苷酸对应前导序列的第28至57个核苷酸,最5'端的81个核苷酸对应mRNA 7的第58至138个核苷酸。No. 8这种结构很可能是由基因组RNA和亚基因组RNA之间的RNA-RNA重组产生的。基因M和N之间的基因间序列中的核苷酸插入导致两个共有序列被111个核苷酸隔开。引物延伸分析表明,由上游共有序列起始合成产生的稍大的亚基因组mRNA的量仅为从下游共有序列起始合成的正常大小的mRNA 7的5%。