Hassan Ahmed O, Vemula Sai V, Sharma Anurag, Bangari Dinesh S, Mishra Krishna K, Mittal Suresh K
Department of Comparative Pathobiology, College of Veterinary Medicine, Purdue University, West Lafayette, IN, USA.
Purdue Institute for Immunology, Inflammation and Infectious Diseases, College of Veterinary Medicine, Purdue University, West Lafayette, IN, USA.
J Gen Virol. 2017 Apr;98(4):749-753. doi: 10.1099/jgv.0.000707. Epub 2017 Apr 27.
Bovine adenovirus (AdV) type 3 (BAdV-3) E1 region shares functional homology with E1 of human AdV type C5. Sequence analysis of the BAdV-3 E1 region revealed the presence of a novel 155R ORF that is not observed in other AdVs, on the lower strand antiparallel to a portion of the E1B region. The 155R gene products in BAdV-3-infected cells were identified by Northern blot, reverse transcriptase PCR followed by sequencing and Western blot analysis using the155R-specific antibody. 155R seems to be a late protein and is present in purified BAdV-3 particles. Replication kinetics of BAdV mutants with either one (BAdV/155R/mt1) or two (BAdV/155R/mt2) stop codons in the 155R ORF were comparable to those of BAdV-3, indicating that 155R is not essential for virus replication in cell culture. These results suggest that 155R-deleted BAdV-3 vectors could be generated in a cell line that fully complements BAdV-3 E1 functions.
牛腺病毒3型(BAdV-3)的E1区域与人腺病毒C5型的E1区域具有功能同源性。对BAdV-3 E1区域的序列分析显示,在与E1B区域一部分反向平行的下链上存在一个在其他腺病毒中未观察到的新型155R开放阅读框。通过Northern印迹、逆转录聚合酶链反应(随后进行测序)以及使用155R特异性抗体的蛋白质印迹分析,鉴定了BAdV-3感染细胞中的155R基因产物。155R似乎是一种晚期蛋白,存在于纯化的BAdV-3颗粒中。在155R开放阅读框中带有一个(BAdV/155R/mt1)或两个(BAdV/155R/mt2)终止密码子的BAdV突变体的复制动力学与BAdV-3相当,这表明155R对于细胞培养中的病毒复制并非必需。这些结果表明,可以在完全补充BAdV-3 E1功能的细胞系中产生缺失155R的BAdV-3载体。