Boyle D B
CSIRO, Australian Animal Health Laboratory, Geelong.
Virus Genes. 1992 Aug;6(3):281-90. doi: 10.1007/BF01702566.
A comparison was undertaken of poxvirus promoters in vaccinia and fowlpox virus (FPV) recombinants using the level of beta-galactosidase expressed from the LacZ gene as a measure of promoter function. In this study a comparison was made of the vaccinia virus promoters, P 7.5 and P L11, the major late promoter of cowpox virus, P CPX (expressing the abundant inclusion body protein), and the FPV promoters, P E/L and P L. In vaccinia virus recombinants the FPV P E/L promoter expressed one-third to one-half the level of beta-galactosidase expressed by the P L11 promoter. In comparison with the P 7.5 promoter, the FPV P E/L promoter expressed four to five times the level of beta-galactosidase. In FPV recombinants beta-galactosidase activity expressed was equal for the P E/L and P CPX promoters. Levels expressed by P L11 and P L were one-half and one-fifth that level, respectively. The temporal regulation of the promoters was maintained in both vaccinia virus and FPV recombinants. The P E/L promoter of FPV has the TAAATG sequence characteristic of late poxvirus promoters at the transcription initiation site. In an attempt to enhance the utility of this promoter for the expression of foreign genes in FPV and vaccinia virus recombinants, the effect upon promoter function of changing the G of the ATG to A, T, or C was determined using transient expression assays with vaccinia virus. Substitution of A, T, or C for the G abolished promoter function. Because of its early/late function, the level of expression and the presence of the oppositely oriented late P L promoter, the FPV P E/L promoter will be valuable for the expression of foreign genes in poxvirus recombinants.
利用从LacZ基因表达的β-半乳糖苷酶水平作为启动子功能的衡量指标，对痘苗病毒和禽痘病毒（FPV）重组体中的痘病毒启动子进行了比较。在本研究中，比较了痘苗病毒启动子P 7.5和P L11、牛痘病毒的主要晚期启动子P CPX（表达丰富的包涵体蛋白）以及FPV启动子P E/L和P L。在痘苗病毒重组体中，FPV的P E/L启动子表达的β-半乳糖苷酶水平是P L11启动子的三分之一到二分之一。与P 7.5启动子相比，FPV的P E/L启动子表达的β-半乳糖苷酶水平高四到五倍。在FPV重组体中，P E/L和P CPX启动子表达的β-半乳糖苷酶活性相当。P L11和P L表达的水平分别是该水平的二分之一和五分之一。启动子的时间调控在痘苗病毒和FPV重组体中均得以维持。FPV的P E/L启动子在转录起始位点具有晚期痘病毒启动子特有的TAAATG序列。为了增强该启动子在FPV和痘苗病毒重组体中表达外源基因的效用，通过痘苗病毒的瞬时表达试验，确定了将ATG中的G替换为A、T或C对启动子功能的影响。用A、T或C替换G会消除启动子功能。由于其早期/晚期功能、表达水平以及反向排列的晚期P L启动子的存在，FPV的P E/L启动子对于在痘病毒重组体中表达外源基因将具有重要价值。
