Pallister J, Wright P J, Sheppard M
Commonwealth Scientific and Industrial Research Organisation Division of Animal Health, Parkville, Victoria, Australia.
J Virol. 1996 Aug;70(8):5115-22. doi: 10.1128/JVI.70.8.5115-5122.1996.
Intertypic recombinant fowl adenoviruses (FAVs) were generated to determine regions of the viral genome involved in virulence. Recombinants were produced with two serotype 8 FAVs, mildly virulent CFA 3 and hypervirulent CFA 40. Restriction endonuclease fragments from the genomes of the two FAVs were used to transfect primary chicken kidney cells. Virulence testing of these recombinants located the region responsible for differences in virulence to an 8.4-kb fragment of the genome located between kb 26.6 and 35.0. According to data available for a serotype 10 FAV that had been partially characterized in the laboratory, this segment of the genome contained three genes of known identity (100K, 33K, and pVIII) and a region between kb 31 and 35 with unknown coding potential (although this information subsequently became available for a serotype 1 FAV, CELO). Therefore, the region between kb 30.5 and 34.5 was sequenced. The results revealed that the unknown region encoded a fiber gene on the right strand and several small open reading frames of unknown identity on the left strand. Further recombinant viruses containing defined exchanges within the 4-kb fragment were constructed, and virulence testing of these viruses indicated that the fiber was responsible for differences in virulence for CFA 40 and CFA 3.
构建了不同血清型重组禽腺病毒(FAV)以确定病毒基因组中与毒力相关的区域。使用两种8型FAV,即低毒力的CFA 3和高毒力的CFA 40产生重组体。来自这两种FAV基因组的限制性内切酶片段用于转染原代鸡肾细胞。对这些重组体的毒力测试将导致毒力差异的区域定位到基因组中位于26.6 kb至35.0 kb之间的一个8.4 kb片段。根据实验室中已部分鉴定的10型FAV的现有数据,该基因组片段包含三个已知身份的基因(100K、33K和pVIII)以及一个位于31 kb至35 kb之间具有未知编码潜能的区域(尽管随后获得了1型FAV CELO的该信息)。因此,对30.5 kb至34.5 kb之间的区域进行了测序。结果显示,未知区域在正义链上编码一个纤维基因,在反义链上编码几个身份未知的小开放阅读框。构建了在4 kb片段内包含特定交换的进一步重组病毒,对这些病毒的毒力测试表明,纤维是CFA 40和CFA 3毒力差异的原因。