Binns M M, Britton B S, Mason C, Boursnell M E
AFRC Institute for Animal Health, Houghton Laboratory, Huntingdon, Cambridgeshire, U.K.
J Gen Virol. 1990 Dec;71 ( Pt 12):2873-81. doi: 10.1099/0022-1317-71-12-2873.
The DNA sequence of the fowlpox virus genome corresponding to the vaccinia virus D6 to A1 region has been determined. Translation of this sequence reveals fowlpoxvirus gene homologues corresponding to the D6, D7, D9, D10, D11, D12, D13 and A1 genes of vaccinia virus. In contrast, no gene homologue for the non-essential vaccinia virus D8 gene was present in fowlpox virus. Instead, a gene transcribed from the opposite strand to the vaccinia virus D8 gene showing no homology to any previously sequenced poxvirus gene was present. The amino terminus of the fowlpox virus D9 homologue had undergone substantial changes, including frameshifts which would be predicted to inactivate the gene. Insertion of a gene cartridge composed of the vaccinia virus p7.5 promoter and the lacZ gene into the fowlpox virus D8, D9 and D10 genes in vitro, followed by recombination into fowlpox virus, was carried out. Stable insertion mutants with the correct genotype were obtained for D8 and D9 which, when tested in chickens did not appear to have been attenuated. No stable insertion mutants were obtained for D10, indicating that this gene probably encodes a function which is essential for virus replication. The D8 and D9 genes of fowlpox virus represent useful insertion sites for the construction of recombinant fowlpox virus vaccines.
已确定与痘苗病毒D6至A1区域相对应的禽痘病毒基因组的DNA序列。对该序列进行翻译后发现,禽痘病毒存在与痘苗病毒D6、D7、D9、D10、D11、D12、D13和A1基因相对应的基因同源物。相比之下,禽痘病毒中不存在痘苗病毒非必需基因D8的基因同源物。相反,存在一个从与痘苗病毒D8基因相反的链转录的基因,该基因与任何先前测序的痘病毒基因均无同源性。禽痘病毒D9同源物的氨基末端发生了重大变化,包括移码,预计这会使该基因失活。在体外将由痘苗病毒p7.5启动子和lacZ基因组成的基因盒插入禽痘病毒D8、D9和D10基因中,随后重组到禽痘病毒中。对于D8和D9,获得了具有正确基因型的稳定插入突变体,在鸡中进行测试时,这些突变体似乎并未减毒。未获得D10的稳定插入突变体,这表明该基因可能编码病毒复制所必需的功能。禽痘病毒的D8和D9基因是构建重组禽痘病毒疫苗的有用插入位点。