Kochneva G V, Babkina I N, Lupan T A, Grazhdantseva A A, Iudin P V, Sivolobova G F, Shvalov A N, Popov E G, Babkin I V, Netesov S V, Chumakov P M
Mol Biol (Mosk). 2013 Sep-Oct;47(5):842-52.
Chicken anemia virus gene encoding apoptin, a selective killer of cancer cells was synthesized and inserted into vaccinia virus (strain L-IVP) genome. The insertion has replaced major part of the viral C11R gene encoding viral growth factor (VGF), which is important for the virulence. The recombinant virus VVdGF-ApoS24/2 was obtained through the transient dominant selection technique with the use of puromycin resistance gene as the selective marker. The expression apoptin gene from a synthetic early-late promoter of vaccinia virus effectively provides accumulation of the protein in the cells infected with the VVdGF-ApoS24/2 virus. Despite the presence of virus growth factor signal peptide at apoptin N-terminal secretion of the recombinant protein into culture medium did not occur. The recombinant virus VVdGF-ApoS24/2 was found to have a significantly greater selective lyticactivity on human cancer cell lines (A549, A431, U87MG, RD and MCF7) as compared with the parent strain L-IVP and its variant VVdGF2/6 with the deletion of the C11R gene. The results suggest that the use of apoptin represents a promising approach for improving the natural anticancer activities of vaccinia virus.
编码凋亡素（一种癌细胞选择性杀伤因子）的鸡贫血病毒基因被合成，并插入痘苗病毒（L-IVP株）基因组。该插入取代了编码病毒生长因子（VGF）的病毒C11R基因的大部分，VGF对病毒毒力很重要。利用嘌呤霉素抗性基因作为选择标记，通过瞬时显性选择技术获得了重组病毒VVdGF-ApoS24/2。来自痘苗病毒合成的早晚期启动子的凋亡素基因表达有效地使该蛋白在感染VVdGF-ApoS24/2病毒的细胞中积累。尽管凋亡素N端存在病毒生长因子信号肽，但重组蛋白并未分泌到培养基中。与亲本株L-IVP及其缺失C11R基因的变体VVdGF2/6相比，发现重组病毒VVdGF-ApoS24/2对人癌细胞系（A549、A431、U87MG、RD和MCF7）具有显著更高的选择性裂解活性。结果表明，使用凋亡素是提高痘苗病毒天然抗癌活性的一种有前景的方法。
