Loparev V N, Chernos V I
Mol Gen Mikrobiol Virusol. 1987 Nov(11):34-9.
The recombinant plasmids pVL1 and pVL2 were constructed for insertion and expression of alien genetic information in HindIII-F fragment of vaccinia virus DNA under the control of the strong early-late promoter of the protein 7.5. The late promoter of the main late protein 11K of vaccinia virus was cloned. These as well as other vector plasmids have been used to express the procaryotic beta-galactosidase gene. Functional activity of the genetic engineering constructions was estimated by transitory expression of beta-galactosidase after plasmid DNA transfection into the chicken fibroblasts embryo culture infected with vaccinia virus. The promoters of the genes for 7.5K and 11K proteins permitted the high level of beta-galactosidase expression. Using of the early promoter of the central part of HindIII-F fragment DNA from vaccinia virus was less efficient for expression of the enzyme.
构建了重组质粒pVL1和pVL2，用于在痘苗病毒DNA的HindIII-F片段中插入和表达外源遗传信息，该过程受蛋白质7.5强早期-晚期启动子的控制。克隆了痘苗病毒主要晚期蛋白11K的晚期启动子。这些质粒以及其他载体质粒已被用于表达原核β-半乳糖苷酶基因。通过将质粒DNA转染到感染痘苗病毒的鸡胚成纤维细胞培养物中后β-半乳糖苷酶的瞬时表达，评估了基因工程构建体的功能活性。7.5K和11K蛋白基因的启动子允许高水平的β-半乳糖苷酶表达。使用痘苗病毒HindIII-F片段DNA中部的早期启动子表达该酶的效率较低。
