用杆状病毒为载体表达Epstein-Barr病毒膜抗原

Baculovirus expression of membrane antigen of Epstein-Barr virus and immunogenic studies of the expressed glycoproteins

用杆状病毒转染载体质粒 pVL941lacZ(含大肠杆菌β-半乳糖苷酶)构建切去3′端穿膜序列Epstein—Barr 病毒(EBV)膜抗原(GPTR)的质粒pVL941 lac GPTR。将 pVL941 lac GPTR DNA 转染昆虫病毒感染的 Sf9细胞,在显色剂 X-gal 存在下筛选无多角体的蓝色蚀斑,经过6次纯化,获得含EBV 膜抗原的重组杆状病毒。1 材料来源 pVL941 lac

A recombinant transfer vector plasmia pv1941LacGPTR carrying Epstein-Barr virus(EBV)membrane antigen (GPTR)gene and LacZ was constructed and was used to obtain recombinant AcNPV after transfection of Sf9 cells infected with AcNPV and selection of occlusion-negative dark blue plagues with the chromogenic dye X-gal. Sf9 cells infected with reconbinant AcNPV were examined by the restriction eenzyme analysis and southernblot.The re- sult showed that the GPTR gene was correctly inserted into recombinant baculovirus DNA adjacent downstream of the polyhedrosis in promotor gene.The GPTR protein expressed were found to be able to bind specifically to the serum of an NPC patient by dot-immunblot assay.Western blot showed that the secreted protein had the same molecular weight as those of nature gp340/220,340KD and 220KD.The truncated by gp340/220 in culture supernatant was concertrated by polyethylene glycol-6000.Immunization test in mice of GPTR expressed in Sf9 cells infected with recombinant AcNPV.