粘虫颗粒体病毒增效因子的基因定位

参考粉纹夜蛾Trichoplusia ni 颗粒体病毒增强因子的基因序列，设计PCR引物，用PCR反应扩增出特异性产物。用EcoRⅠ、BamHⅠ双酶酶切处理PCR反应产物，然后克隆到质粒pUC19中，构建重组质粒pUC19-SF；对重组质粒pUC19-SF中的外源片段测序，结果证明PCR扩增产物是粘虫颗粒体病毒PuGV-Ps增效因子基因的一段序列。重组质粒pUC19-SF的插入片段标记为探针，通过Southern杂交将增效因子基因定位于PuGV-Ps病毒基因组的多种酶切片段上。

Location of the gene encoding synergistic factor of the Pseudaletia unipuncta granulosis virus

Using the nucleotide sequence of the VEF (viral enhancing factor) gene of Trichoplusia ni granulosis virus for reference, we designed a primer for PCR and amplified a specific fragment by PCR reaction. A recombinant plasmid pUC19-SF was constructed by cloning the PCR product which was treated with EcoRⅠ, BamHⅠ into pUC19. The PCR product was proven to be a fragment of the gene encoding the synergistic factor of Pseudaletia unipuncta granulosis virus (PuGV-Ps) by DNA sequencing. After labeling this fragment as a probe, the gene encoding of the synergistic factor was located in many restrictive fragments of PuGV-Ps genome by Southern blot hybridization.