title: Construction of the ie1-Bacmid expression system and its use to express EGFP and BmAGO2 in BmN cells.
abstract: The presently available expression tools and vectors (e.g., eukaryotic expression vectors and the adenovirus expression system) for studying the functional genes in Bombyx mori are insufficient. The baculovirus expression system is only used as a protein production tool; therefore, recombinant proteins expressed by B. mori using the baculovirus expression system equipped with a polyhedrin promoter cannot be used for in vivo research applications. In this work, we constructed and screened a eukaryotic expression vector for silkworm cells The EGFP and B. mori Argonaute2 proteins were found to be efficiently expressed using the screened pIEx-1 vector with the FuGENE 6 transfection reagent. Additionally, we constructed a novel nucleopolyhedrovirus ie1-Bacmid expression system for the production of recombinant protein; we then used the system to highly express the EGFP and B. mori Argonaute2 proteins. In this system, the protein of interest can be efficiently expressed 13 h after infection by controlling the B. mori nucleopolyhedrovirus immediate early ie1 promoter. The ie1-Bacmid system provides a powerful "adenovirus-like" expression tool; not only can the tool be used to study baculovirus molecular biology for the silkworm but it is also useful in other research applications as well, such as the study of gene functions involved in cellular physiological processes.
authors: Institute of Biochemistry, College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou, 310018, China.
authors: Fang
authors: F
authors: Zhou
authors: Zhen
authors: Z
authors: Gao
authors: Zhengbing
authors: Z
authors: Lv
authors: Jian
authors: J
authors: Chen
authors: Yeting
authors: Y
authors: Hong
authors: Wei
authors: W
authors: Yu
authors: Dan
authors: D
authors: Wang
authors: Caiying
authors: C
authors: Jiang
authors: Xiangfu
authors: X
authors: Wu
authors: Yaozhou
authors: Y
authors: Zhang
authors: Zuoming
authors: Z
authors: Nie
mesh_terms: D000818:Animals; D060565:Argonaute Proteins / Q000235:genetics / Q000378:metabolism*; D012831:Bombyx / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D008954:Models, Biological; D011401:Promoter Regions, Genetic / Q000235:genetics
chemical_list: D060565:Argonaute Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins