Hofmann M A, Brian D A
Department of Microbiology, University of Tennessee, Knoxville 37996-0845.
J Virol. 1991 Nov;65(11):6331-3. doi: 10.1128/JVI.65.11.6331-6333.1991.
A radiolabeled oligodeoxynucleotide primer that anneals near the common 5' end of bovine coronavirus minus-strand RNAs was extended with reverse transcriptase, and a major product suggesting poly(U) tracts of 8 to 20 nucleotides was found. The extended primer molecules were ligated head to tail, amplified by the polymerase chain reaction, cloned, and sequenced, and poly(U) tracts of 9 to 26 nucleotides were found. Poly(A) tails of 100 to 130 nucleotides on the 3' end of coronavirus plus-strand mRNAs and genome must, therefore, be generated by a mechanism that uses only a short poly(U) template. This pattern contrasts with that of other cytoplasmic, polyadenylated, plus-strand animal RNA viruses which utilize a full-length poly(U) template for poly(A) synthesis.
一种放射性标记的寡脱氧核苷酸引物与牛冠状病毒负链RNA的共同5'端附近退火,用逆转录酶进行延伸,发现了一个主要产物,提示存在8至20个核苷酸的聚(U)序列。延伸的引物分子头尾相连进行连接,通过聚合酶链反应进行扩增、克隆和测序,发现了9至26个核苷酸的聚(U)序列。因此,冠状病毒正链mRNA和基因组3'端100至130个核苷酸的聚(A)尾必定是由一种仅使用短聚(U)模板的机制产生的。这种模式与其他利用全长聚(U)模板进行聚(A)合成的细胞质、多聚腺苷酸化、正链动物RNA病毒的模式形成对比。