Boyle D B, Coupar B E, Gibbs A J, Seigman L J, Both G W
Virology. 1987 Feb;156(2):355-65. doi: 10.1016/0042-6822(87)90415-6.
The thymidine kinase (TK) gene of fowlpox virus (FPV) is located in a 2.2-kb HindIII-ClaI fragment derived from a 5.5-kb EcoR1 fragment of the FPV genome. The TK gene was mapped to the region of a 700-bp XbaI fragment contained within this HindIII-ClaI fragment. Nucleotide sequence analysis of this region revealed an open reading frame of 183 codons. Identification of this region as the FPV TK gene was confirmed by its homology with the vaccinia virus TK at both the nucleotide and amino acid levels. The derived FPV TK polypeptide has a calculated molecular weight of 20,380 and is six amino acids larger than the vaccinia virus TK gene product. We have reported previously that the FPV TK gene operates in vaccinia virus without the requirement for a vaccinia virus promoter. The sequence homologies between the two TK promoters substantiated this observation. Northern blot analysis of RNAs from cells infected with a vaccinia virus recombinant expressing the FPV TK gene showed major (700 nucleotide) and minor (1000 nucleotide) transcripts from the FPV TK gene. The deduced amino acid sequence of the FPV TK has significant homology with the TKs from chicken, man, and three other poxviruses, but shows no homology with herpes simplex virus TK. Comparisons of the homologous sequences indicated that the "core" of the enzyme has probably evolved in poxviruses four times as quickly as in vertebrates. Characterization of the FPV TK gene may facilitate the construction of recombinant FPVs as vehicles for the delivery of vaccine antigens to poultry and other avian species.
禽痘病毒(FPV)的胸苷激酶(TK)基因位于一个2.2kb的HindIII-ClaI片段中,该片段源自FPV基因组的一个5.5kb EcoR1片段。TK基因被定位到这个HindIII-ClaI片段内一个700bp的XbaI片段区域。对该区域的核苷酸序列分析揭示了一个由183个密码子组成的开放阅读框。通过在核苷酸和氨基酸水平上与痘苗病毒TK的同源性,证实了该区域为FPV TK基因。推导的FPV TK多肽的计算分子量为20380,比痘苗病毒TK基因产物大六个氨基酸。我们之前报道过,FPV TK基因在痘苗病毒中起作用,无需痘苗病毒启动子。两个TK启动子之间的序列同源性证实了这一观察结果。对感染表达FPV TK基因的痘苗病毒重组体的细胞中的RNA进行Northern印迹分析,显示出来自FPV TK基因的主要(700个核苷酸)和次要(1000个核苷酸)转录本。推导的FPV TK氨基酸序列与鸡、人及其他三种痘病毒的TK有显著同源性,但与单纯疱疹病毒TK无同源性。同源序列比较表明,该酶的“核心”在痘病毒中的进化速度可能是脊椎动物中的四倍。对FPV TK基因的特性分析可能有助于构建重组FPV,作为向家禽和其他禽类递送疫苗抗原的载体。