Wu C H, Wang C C, Yang H M, Wang A L
Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan.
Gene. 1995 May 26;158(1):129-31. doi: 10.1016/0378-1119(95)00092-k.
The linear double-stranded RNA (dsRNA) genome of giardiavirus (GLV) was estimated to be 6100 nucleotides (nt) [Wang et al., Proc. Natl. Acad. Sci. USA 90 (1993) 8585-8599]. As the 5'- and 3'-untranslated regions (UTR) of viral genomes are known to contain critical information for viral replication, we reexamine the sequences at all four termini of GLV dsRNA by (i) direct RNA sequencing with RT, (ii) tailing GLV dsRNA with UTP or CTP in addition to ATP in 3' rapid amplification of cDNA ends (RACE) and (iii) adding poly(dG) to the products of primer extension in 5'-RACE. The results confirmed the reported sequence for the 5'-terminus of the GLV sense strand RNA, but uncovered an additional 177 nt at the 3'-terminus. The new study also showed conclusively that there are no protruding overhangs at either terminus of GLV dsRNA.
贾第虫病毒(GLV)的线性双链RNA(dsRNA)基因组估计有6100个核苷酸(nt)[Wang等人,《美国国家科学院院刊》90(1993)8585 - 8599]。由于已知病毒基因组的5'和3'非翻译区(UTR)包含病毒复制的关键信息,我们通过以下方法重新检查了GLV dsRNA所有四个末端的序列:(i)用逆转录酶进行直接RNA测序,(ii)在3' cDNA末端快速扩增(RACE)中除了ATP外还用UTP或CTP对GLV dsRNA进行加尾,以及(iii)在5'-RACE中对引物延伸产物添加聚(dG)。结果证实了已报道的GLV正义链RNA 5'末端的序列,但在3'末端发现了另外177个nt。这项新研究还确凿地表明,GLV dsRNA的任何一个末端都没有突出的悬端。