Dryga S A, Sviatchenko V A, Mikriukova T P, Fursova E Iu, Kiselev N N, Goncharov A M, Frolov I V, Kolykhalov A A, Agapov E V, Netesov S V
Vopr Virusol. 1996 May-Jun;41(3):100-4.
Venezuelan equine encephalomyelitis (VEE) virus-based vectors for expression of heterologous genes have been constructed using full-length cDNA copy of the interstrain recombinant virus (Trinidad Donkey and 230 strain). Gene cassettes carrying the subgenomic mRNA promoter of VEE virus and the preS2-S gene of hepatitis B virus (HBV) were inserted before or after the genes of structural viral proteins. Live virus stocks were obtained by transfection of chick embryo fibroblasts with in vitro transcribed full-length RNA. Insertions of gene cassettes before the structural region resulted in expression of HBsAg (VEHB-25 and VEHB-361 viruses), whereas insertions in the 3' region did not. Recombinant virus VEHB-25 expressed HBsAg during 5 passages in Vero cells. VEHB-25 stimulated immune response to HBsAg and was less virulent than the parental virus.
已利用株间重组病毒(特立尼达驴和230株)的全长cDNA拷贝构建了用于表达异源基因的基于委内瑞拉马脑脊髓炎(VEE)病毒的载体。携带VEE病毒亚基因组mRNA启动子和乙型肝炎病毒(HBV)前S2 - S基因的基因盒被插入病毒结构蛋白基因之前或之后。通过用体外转录的全长RNA转染鸡胚成纤维细胞获得活病毒储备。在结构区域之前插入基因盒导致HBsAg表达(VEHB - 25和VEHB - 361病毒),而在3'区域插入则不表达。重组病毒VEHB - 25在Vero细胞中传代5次期间表达HBsAg。VEHB - 25刺激了对HBsAg的免疫反应,并且比亲本病毒的毒性小。