Soldevila A I, Webb B A
Department of Entomology, University of Kentucky, Lexington, KY 40546-0091, USA.
J Gen Virol. 1996 Jul;77 ( Pt 7):1379-88. doi: 10.1099/0022-1317-77-7-1379.
We have evaluated the use of baculoviruses to deliver Campoletis sonorensis polydnavirus (CsPDV) genomic DNA into lepidopteran larvae to facilitate the identification of functional CsPDV genes. Genomic fragments consisting of regulatory (promoter) and coding sequences for two CsPDV genes (VHv1.1 and WHv1.6) were used to generate CsPDV-baculovirus recombinants and evaluate the expression of genes under the regulation of the CsPDV promoters. Northern blot and primer extension studies established that CsPDV genes were expressed under the control of their own promoters in these CsPDV-baculovirus recombinants. Transcripts were detected as early as 4 h post-infection indicating that temporal activity of CsPDV promoters was retained. The VHv1.1 gene product as expressed from CsPDV-baculovirus recombinants was identical in size and in functional properties to that produced in CsPDV-infected insects. CsPDV-baculovirus recombinants may be useful for the screening and characterization of polydnavirus genes with functional activities that can only be evaluated in insect larvae.
我们评估了利用杆状病毒将索诺拉 Campoletis 多角体病毒（CsPDV）基因组 DNA 导入鳞翅目幼虫，以促进 CsPDV 功能基因的鉴定。由两个 CsPDV 基因（VHv1.1 和 WHv1.6）的调控（启动子）和编码序列组成的基因组片段用于产生 CsPDV - 杆状病毒重组体，并评估 CsPDV 启动子调控下基因的表达。Northern 印迹和引物延伸研究表明，在这些 CsPDV - 杆状病毒重组体中，CsPDV 基因在其自身启动子的控制下表达。早在感染后 4 小时就检测到转录本，表明 CsPDV 启动子的时间活性得以保留。从 CsPDV - 杆状病毒重组体表达的 VHv1.1 基因产物在大小和功能特性上与 CsPDV 感染昆虫中产生的相同。CsPDV - 杆状病毒重组体可能有助于筛选和表征具有只能在昆虫幼虫中评估的功能活性的多角体病毒基因。
