Massung R F, Moyer R W
Department of Immunology and Medical Microbiology, College of Medicine, J. Hills Miller Health Center, University of Florida, Gainesville 32610.
J Virol. 1990 May;64(5):2280-9. doi: 10.1128/JVI.64.5.2280-2289.1990.
We have examined the feasibility of using Xenopus laevis oocytes microinjected with rabbit poxvirus as a system to study poxvirus gene expression. The injection of either intact virus or subviral cores resulted in accurate synthesis of viral proteins. This expression was dependent on the multiplicity of injected virus, with the optimal injected dose being equivalent to approximately 300 PFU per oocyte. Extensive viral gene expression including late viral protein synthesis was observed when intact virions were microinjected into the oocyte. However, the injection of subviral cores resulted in only early protein synthesis. When oocytes were injected with a mixture of subviral cores and the nonionic detergent-soluble fraction was removed from virus during the preparation of cores, both early and late viral proteins were synthesized. Therefore, the detergent-soluble fraction appears to contain a factor(s) required for the transition from early to late gene expression.
我们研究了将注射了兔痘病毒的非洲爪蟾卵母细胞作为研究痘病毒基因表达系统的可行性。注射完整病毒或亚病毒核心均可导致病毒蛋白的准确合成。这种表达依赖于注射病毒的感染复数,最佳注射剂量约为每个卵母细胞300个蚀斑形成单位(PFU)。当将完整病毒粒子显微注射到卵母细胞中时,观察到广泛的病毒基因表达,包括晚期病毒蛋白合成。然而,注射亚病毒核心仅导致早期蛋白合成。当用亚病毒核心混合物注射卵母细胞,且在制备核心过程中从病毒中去除非离子去污剂可溶部分时,早期和晚期病毒蛋白均会合成。因此,去污剂可溶部分似乎含有从早期基因表达过渡到晚期基因表达所需的一种或多种因子。