[Construction and characterization of a novel baculovirus vector based on the White spot syndrome virus immediately-early promoter].
OBJECTIVE
To construct a universal baculovirus vector for efficient gene expression in both invertebrate and vertebrate cell lines.


METHODS
Using the Bac-To-Bac system, we genetically engineered the immediately-early 1 gene promoter (ie1 promoter) from White spot syndrome virus into a baculovirus vector that was pseudotyped with Vesicular stomatitis virus glycoprotein (VSV G). We placed the enhanced green fluorescent protein (EGFP) gene under the control of iel promoter in the baculovirus vector to get the reporter recombinant baculovirus, vAc-G-EGFP. We tested the reporter EGFP gene expression in tested cell lines through virus infection or transduction experiments using direct fluorescence microscopy and Western blot analysis.


RESULTS
Under the control of ie1 promoter, vAc-G-EGFP could efficiently express the EGFP reporter gene in both invertebrate and vertebrate cells. The steady-state expression level of EGFP in vertebrate cell lines were different from that in invertebrate Sf9 cells as reflected by Western blot assays.


CONCLUSIONS
The iel promoter-based and VSV G-pseudotyped baculovirus vector presents a unique and effective tool to express target genes simultaneously in various cell systems; the novel baculovirus-mediated gene expression system developed in this study has the potential to be widely used in both basic and applied research.
Lu
Liqun
L
Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, Ministry of Education, Shanghai Ocean University, Shanghai 201306, China. lqlv@shou.edu.cn
Xu
Hongxu
H
Wang
Hao
H
D000818:Animals; D016367:Baculoviridae / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D015870:Gene Expression; D017930:Genes, Reporter; D005818:Genetic Engineering; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D018411:Spodoptera; D046848:White spot syndrome virus 1 / Q000235:genetics*
C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins