Page K W, Mawditt K L, Britton P
AFRC Institute for Animal Health, Compton Laboratory, Newbury, Berkshire, U.K.
J Gen Virol. 1991 Mar;72 ( Pt 3):579-87. doi: 10.1099/0022-1317-72-3-579.
Analysis of porcine transmissible gastroenteritis virus (TGEV) and porcine respiratory coronavirus (PRCV) mRNA species indicated a deletion in mRNA 3 of PRCV. Polymerase chain reaction (PCR) was used to clone the 5' end of mRNA 3 from PRCV for comparison with the equivalent region in TGEV. Small deletions were observed within and around the PRCV sequence equivalent to the putative open reading frame (ORF) ORF-3a identified in TGEV. The potential RNA polymerase-leader complex binding site (leader RNA binding site), ACTAAAC, found upstream of ORF-3a in TGEV, was absent from the PRCV genome but a potential site was found in the PRCV genome upstream of a gene equivalent to TGEV ORF-3b. PCR analysis, using primers corresponding to sequences within the ORF-3b gene and the leader RNA sequence, confirmed that the leader RNA binding site was upstream of a gene equivalent to TGEV ORF-3b on PRCV mRNA 3 but upstream of ORF-3a on TGEV mRNA 3. The presence of the new leader RNA binding site would be responsible for generating the smaller mRNA 3 species found in PRCV-infected cells.
对猪传染性胃肠炎病毒(TGEV)和猪呼吸道冠状病毒(PRCV)mRNA种类的分析表明,PRCV的mRNA 3存在缺失。采用聚合酶链反应(PCR)从PRCV中克隆mRNA 3的5'端,以便与TGEV中的相应区域进行比较。在与TGEV中鉴定的推定开放阅读框(ORF)ORF-3a等效的PRCV序列内部及周围观察到小的缺失。TGEV中位于ORF-3a上游的潜在RNA聚合酶-前导序列复合物结合位点(前导RNA结合位点)ACTAAAC在PRCV基因组中不存在,但在与TGEV ORF-3b等效的基因上游的PRCV基因组中发现了一个潜在位点。使用与ORF-3b基因内序列和前导RNA序列对应的引物进行的PCR分析证实,前导RNA结合位点在PRCV mRNA 3上与TGEV ORF-3b等效的基因上游,但在TGEV mRNA 3上位于ORF-3a上游。新前导RNA结合位点的存在将导致在PRCV感染细胞中发现较小的mRNA 3种类。